PB-iETV2

LOCUS       PB-iETV2      8506 bp DNA     circular SYN 29-APR-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8506)
  AUTHORS   Amane
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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..1811
                     /label=653-ETV2-F
     regulatory      1789..1798
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1795..2823
                     /codon_start=1
                     /label=ETV2
                     /translation="MDLWNWDEASPQEVPPGNKLAGLEGAKLGFCFPDLALQGDTPTAT
                     AETCWKGTSSSLASFPQLDWGSALLHPEVPWGAEPDSQALPWSGDWTDMACTAWDSWSG
                     ASQTLGPAPLGPGPIPAAGSEGAAGQNCVPVAGEATSWSRAQAAGSNTSWDCSVGPDGD
                     TYWGSGLGGEPRTDCTISWGGPAGPDCTTSWNPGLHAGGTTSLKRYQSSALTVCSEPSP
                     QSDRASLARCPKTNHRGPIQLWQFLLELLHDGARSSCIRWTGNSREFQLCDPKEVARLW
                     GERKRKPGMNYEKLSRGLRYYYRRDIVRKSGGRKYTYRFGGRVPSLAYPDCAGGGRGAE
                     TQ"
     primer_bind     complement(2803..2838)
                     /label=653-ETV2-R
     polyA_signal    2836..3043
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3120..3331
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             3646..4389
                     /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             4390..4443
                     /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             4444..5043
                     /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    5076..5197
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    complement(5686..5998)
                     /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(6485..6501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6509..6525
                     /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(6533..6563)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6578..6599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6887..7475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7646..8506)
                     /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 caccatggat
     1801 ttgtggaatt gggatgaggc atcacctcaa gaggttccgc ctggtaacaa acttgcaggg
     1861 ctcgaaggag ccaaattagg cttctgtttc cctgatctgg cactccaagg ggacacgccg
     1921 acagcgacag cagagacatg ctggaaaggt acaagctcat ccctggcaag cttcccacag
     1981 ctggactggg gctccgcgtt actgcaccca gaagttccat ggggggcgga gcccgactct
     2041 caggctcttc cgtggtccgg ggactggaca gacatggcgt gcacagcctg ggactcttgg
     2101 agcggcgcct cgcagaccct gggccccgcc cctctcggcc cgggccccat ccccgccgcc
     2161 ggctccgaag gcgccgcggg ccagaactgc gtccccgtgg cgggagaggc cacctcgtgg
     2221 tcgcgcgccc aggccgccgg gagcaacacc agctgggact gttctgtggg gcccgacggc
     2281 gatacctact ggggcagtgg cctgggcggg gagccgcgca cggactgtac catttcgtgg
     2341 ggcgggcccg cgggcccgga ctgtaccacc tcctggaacc cggggctgca tgcgggtggc
     2401 accacctctt tgaagcggta ccagagctca gctctcaccg tttgctccga accgagcccg
     2461 cagtcggacc gtgccagttt ggctcgatgc cccaaaacta accaccgagg tcccattcag
     2521 ctgtggcagt tcctcctgga gctgctccac gacggggcgc gtagcagctg catccgttgg
     2581 actggcaaca gccgcgagtt ccagctgtgc gaccccaaag aggtggctcg gctgtggggc
     2641 gagcgcaaga gaaagccggg catgaattac gagaagctga gccggggcct tcgctactac
     2701 tatcgccgcg acatcgtgcg caagagcggg gggcgaaagt acacgtaccg cttcgggggc
     2761 cgcgtgccca gcctagccta tccggactgt gcgggaggcg gacggggagc agagacacaa
     2821 taaggtaccc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2881 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2941 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3001 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctcttcga
     3061 atttaaatcg gatccagctg aatctaagtg gatctgcgat cgctccggtg cccgtcagtg
     3121 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
     3181 gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     3241 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     3301 ttttcgcaac gggtttgccg ccagaacaca gctgaagctt cgaggggctc gcatctctcc
     3361 ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc cggttgagtc gcgttctgcc
     3421 gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc taggtaagtt taaagctcag
     3481 gtcgagaccg ggcctttgtc cggcgctccc ttggagccta cctagactca gccggctctc
     3541 cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt gtttcgtttt ctgttctgcg
     3601 ccgttacaga tctaagctgt gaccggcgcc tactctagag ccgccatgtc tagactggac
     3661 aagagcaaag tcataaactc tgctctggaa ttactcaatg gagtcggtat cgaaggcctg
     3721 acgacaagga aactcgctca aaagctggga gttgagcagc ctaccctgta ctggcacgtg
     3781 aagaacaagc gggccctgct cgatgccctg ccaatcgaga tgctggacag gcatcatacc
     3841 cactcctgcc ccctggaagg cgagtcatgg caagactttc tgcggaacaa cgccaagtca
     3901 taccgctgtg ctctcctctc acatcgcgac ggggctaaag tgcatctcgg cacccgccca
     3961 acagagaaac agtacgaaac cctggaaaat cagctcgcgt tcctgtgtca gcaaggcttc
     4021 tccctggaga acgcactgta cgctctgtcc gccgtgggcc actttacact gggctgcgta
     4081 ttggaggaac aggagcatca agtagcaaaa gaggaaagag agacacctac caccgattct
     4141 atgcccccac ttctgaaaca agcaattgag ctgttcgacc ggcagggagc cgaacctgcc
     4201 ttccttttcg gcctggaact aatcatatgt ggcctggaga aacagctaaa gtgcgaaagc
     4261 ggcgggccga ccgacgccct tgacgatttt gacttagaca tgctcccagc cgatgccctt
     4321 gacgactttg accttgatat gctgcctgct gacgctcttg acgattttga ccttgacatg
     4381 ctccccgggg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     4441 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     4501 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     4561 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     4621 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     4681 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     4741 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     4801 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     4861 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     4921 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     4981 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5041 tgaaatcaac ctctggatta caaaatttgt gggtaaactt gtttattgca gcttataatg
     5101 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5161 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggaat tgactcaaat
     5221 gatgtcaatt agtctatcag aagctcatct ggtctccctt ccgggggaca agacatccct
     5281 gtttaatatt taaacagcag tgttcccaaa ctgggttctt atatcccttg ctctggtcaa
     5341 ccaggttgca gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac agtggcagcc
     5401 aggtttagcc ccggaattga ctggattcct tttttagggc ccattggtat ggctttttcc
     5461 ccgtatcccc ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg
     5521 atcccgtgcc accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg
     5581 gggggagcgc cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga
     5641 gggacgtaat tacatccctg ggggctttgg gggggggctg tccctgatat ctataacaag
     5701 aaaatatata tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg
     5761 tatgagttaa atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg
     5821 tcgttatagt tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc
     5881 caagcggcga ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga
     5941 gagcaatatt tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaatc
     6001 tagctgcatc aggatcatat cgtcgggtct tttttccggc tcagtcatcg cccaagctgg
     6061 cgctatctgg gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca
     6121 tggaaagagt ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt
     6181 accatgatga ttcgggaagg tgtggccatg cacgccttta acggtgaact gttcgttcag
     6241 gccacctggg ataccagttc gtcgcggctt ttccggacac agttccggat ggtcagcccg
     6301 aagcgcatca gcaacccgaa caataccggc gacagccgga actgccgtgc cggtgtgcag
     6361 attaatgaca gcggtgcggc gctgggatat tacgtcagcg aggacgggta tcctggctgg
     6421 atgccgcaga aatggacatg gataccccgt gagttacccg gcgggcgcgc ttggcgtaat
     6481 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     6541 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     6601 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     6661 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     6721 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     6781 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     6841 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     6901 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     6961 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     7021 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     7081 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     7141 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     7201 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     7261 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     7321 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     7381 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     7441 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     7501 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     7561 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     7621 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     7681 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     7741 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     7801 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     7861 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     7921 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7981 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     8041 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     8101 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     8161 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     8221 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     8281 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     8341 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     8401 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     8461 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcat
//