PB-iNKX3.1-HA

LOCUS       PB-iNKX3.1-HA               8269 bp DNA     circular SYN 02-JUL-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8269)
  AUTHORS   Amane
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8269
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2497..2583
                     /note="/dnas_title=3xHA-TurboID-NLS in pUC-GW-Amp"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    870..1103
                     /label=3' ITR
                     /note="piggyBac 3' inverted terminal repeatApplication 
                     Notes:Recognized by PBase transposase; DNA flanked by 
                     piggyBac 5' ITR and 3' ITR can be transposed by PBase into 
                     TTAA sites.
                     "
     promoter        1385..1763
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    1393..1411
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1429..1447
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1465..1483
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1501..1519
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1537..1555
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1573..1591
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1609..1627
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    1764..1788
                     /label=MCS
     primer_bind     1774..1812
                     /label=665-NKX3-1-F
     misc_feature    1789..1794
                     /label=Kozak seq
     CDS             1795..2496
                     /codon_start=1
                     /label=NKX3.1
                     /translation="MLRVPEPRPGEAKAEGAAPPTPSKPLTSFLIQDILRDGAQRQGGR
                     TSSQRQRDPEPEPEPEPEGGRSRAGAQNDQLSTGPRAAPEEAETLAETEPERHLGSYLL
                     DSENTSGALPRLPQTPKQPQKRSRAAFSHTQVIELERKFSHQKYLSAPERAHLAKNLKL
                     TETQVKIWFQNRRYKTKRKQLSSELGDLEKHSSLPALKEEAFSRASLVSVYNSYPYYPY
                     LYCVGSWSPAFW"
     primer_bind     complement(2477..2496)
                     /label=665-NKX3-1-R
     primer_bind     2482..2520
                     /label=665-HA-F
     CDS             2497..2523
                     /codon_start=1
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2527..2553
                     /codon_start=1
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2557..2583
                     /codon_start=1
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2565..2606)
                     /label=665-HA-R
     polyA_signal    2599..2806
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2883..3094
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3409..4152
                     /codon_start=1
                     /product="modified rtTA protein that binds tightly to 
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /label=Tet-On(R) 3G
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     CDS             4153..4206
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4207..4806
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4839..4960
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(5449..5761)
                     /label=5' ITR
                     /note="piggyBac 5' inverted terminal repeatRecognized by 
                     PBase transposase; DNA flanked by piggyBac 5' ITR and 3' 
                     ITR can be transposed by PBase into TTAA sites.
                     "
     primer_bind     complement(6248..6264)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6272..6288
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6296..6326)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6341..6362
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6650..7238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7409..8269)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtctttcgtc ttcaagaatt
     1381 cctcgagttt actccctatc agtgatagag aacgtatgaa gagtttactc cctatcagtg
     1441 atagagaacg tatgcagact ttactcccta tcagtgatag agaacgtata aggagtttac
     1501 tccctatcag tgatagagaa cgtatgacca gtttactccc tatcagtgat agagaacgta
     1561 tctacagttt actccctatc agtgatagag aacgtatatc cagtttactc cctatcagtg
     1621 atagagaacg tataagcttt aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt
     1681 agtgaaccgt cagatcgcct ggagcaattc cacaacactt ttgtcttata ccaactttcc
     1741 gtaccacttc ctaccctcgt aaaggtctag agctagcggc gcgccgttgc caccatgctc
     1801 agggttccgg agccgcggcc cggggaggcg aaagcggagg gggccgcgcc gccgaccccg
     1861 tccaagccgc tcacgtcctt cctcatccag gacatcctgc gggacggcgc gcagcggcaa
     1921 ggcggccgca cgagcagcca gagacagcgc gacccggagc cggagccaga gccagagcca
     1981 gagggaggac gcagccgcgc cggggcgcag aacgaccagc tgagcaccgg gccccgcgcc
     2041 gcgccggagg aggccgagac gctggcagag accgagccag aaaggcactt ggggtcttat
     2101 ctgttggact ctgaaaacac ttcaggcgcc cttccaaggc ttccccaaac ccctaagcag
     2161 ccgcagaagc gctcccgagc tgccttctcc cacactcagg tgatcgagtt ggagaggaag
     2221 ttcagccatc agaagtacct gtcggcccct gaacgggccc acctggccaa gaacctcaag
     2281 ctcacggaga cccaagtgaa gatatggttc cagaacagac gctataagac taagcgaaag
     2341 cagctctcct cggagctggg agacttggag aagcactcct ctttgccggc cctgaaagag
     2401 gaggccttct cccgggcctc cctggtctcc gtgtataaca gctatcctta ctacccatac
     2461 ctgtactgcg tgggcagctg gagcccagct ttttggtacc cgtatgatgt tccggattac
     2521 gctggctatc cctacgacgt gcccgactat gccgggtacc cctatgacgt cccagactac
     2581 gcatgaggta cccctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2641 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2701 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2761 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctctt
     2821 cgaatttaaa tcggatccag ctgaatctaa gtggatctgc gatcgctccg gtgcccgtca
     2881 gtgggcagag cgcacatcgc ccacagtccc cgagaagttg gggggagggg tcggcaattg
     2941 aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa agtgatgtcg tgtactggct
     3001 ccgccttttt cccgagggtg ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt
     3061 tctttttcgc aacgggtttg ccgccagaac acagctgaag cttcgagggg ctcgcatctc
     3121 tccttcacgc gcccgccgcc ctacctgagg ccgccatcca cgccggttga gtcgcgttct
     3181 gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc gtctaggtaa gtttaaagct
     3241 caggtcgaga ccgggccttt gtccggcgct cccttggagc ctacctagac tcagccggct
     3301 ctccacgctt tgcctgaccc tgcttgctca actctacgtc tttgtttcgt tttctgttct
     3361 gcgccgttac agatctaagc tgtgaccggc gcctactcta gagccgccat gtctagactg
     3421 gacaagagca aagtcataaa ctctgctctg gaattactca atggagtcgg tatcgaaggc
     3481 ctgacgacaa ggaaactcgc tcaaaagctg ggagttgagc agcctaccct gtactggcac
     3541 gtgaagaaca agcgggccct gctcgatgcc ctgccaatcg agatgctgga caggcatcat
     3601 acccactcct gccccctgga aggcgagtca tggcaagact ttctgcggaa caacgccaag
     3661 tcataccgct gtgctctcct ctcacatcgc gacggggcta aagtgcatct cggcacccgc
     3721 ccaacagaga aacagtacga aaccctggaa aatcagctcg cgttcctgtg tcagcaaggc
     3781 ttctccctgg agaacgcact gtacgctctg tccgccgtgg gccactttac actgggctgc
     3841 gtattggagg aacaggagca tcaagtagca aaagaggaaa gagagacacc taccaccgat
     3901 tctatgcccc cacttctgaa acaagcaatt gagctgttcg accggcaggg agccgaacct
     3961 gccttccttt tcggcctgga actaatcata tgtggcctgg agaaacagct aaagtgcgaa
     4021 agcggcgggc cgaccgacgc ccttgacgat tttgacttag acatgctccc agccgatgcc
     4081 cttgacgact ttgaccttga tatgctgcct gctgacgctc ttgacgattt tgaccttgac
     4141 atgctccccg gggagggcag aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc
     4201 ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     4261 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     4321 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     4381 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     4441 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4501 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     4561 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4621 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     4681 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4741 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4801 gcctgaaatc aacctctgga ttacaaaatt tgtgggtaaa cttgtttatt gcagcttata
     4861 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4921 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg aattgactca
     4981 aatgatgtca attagtctat cagaagctca tctggtctcc cttccggggg acaagacatc
     5041 cctgtttaat atttaaacag cagtgttccc aaactgggtt cttatatccc ttgctctggt
     5101 caaccaggtt gcagggtttc ctgtcctcac aggaacgaag tccctaaaga aacagtggca
     5161 gccaggttta gccccggaat tgactggatt ccttttttag ggcccattgg tatggctttt
     5221 tccccgtatc cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa
     5281 gcgatcccgt gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat
     5341 gcggggggag cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg
     5401 ggagggacgt aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac
     5461 aagaaaatat atatataata agttatcacg taagtagaac atgaaataac aatataatta
     5521 tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg
     5581 cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag
     5641 ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag
     5701 agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta
     5761 atctagctgc atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc
     5821 tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg
     5881 gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg
     5941 tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt
     6001 caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc
     6061 ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg
     6121 cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc
     6181 tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt
     6241 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     6301 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     6361 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     6421 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     6481 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     6541 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     6601 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     6661 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6721 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6781 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6841 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6901 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6961 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7021 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7081 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7141 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7201 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7261 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7321 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     7381 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     7441 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     7501 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     7561 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     7621 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     7681 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     7741 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     7801 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     7861 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7921 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7981 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     8041 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     8101 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     8161 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     8221 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcat
//