pDJ685

LOCUS      pDJ685              6628 bp DNA     circular SYN 08-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6628)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    196..226
                     /label=SleepingBeauty-ITR_Lo
                     /label=nonstandard type:  ITR
     misc_feature    392..423
                     /label=SleepingBeauty-ITR_Li
                     /label=nonstandard type:  ITR
     misc_feature    477..1580
                     /label=EF-1a
                     /note="/vntifkey=21"
     promoter        493..1671
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     562..588
                     /label=3-EFS-F
     primer_bind     606..627
                     /label=65-Ef1a-F
     intron          724..1662
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     misc_feature    1730..2303
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     1833..1858
                     /label=222-WKG-1045-F2
     primer_bind     1996..2017
                     /label=104-IRES-Fseq2
     primer_bind     2236..2254
                     /label=100-IRES-Fseq
     CDS             2353..2952
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     2487..2525
                     /label=223-PuroR-F
     polyA_signal    3001..3225
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3230..3251
                     /label=399-1157-F
     misc_feature    3322..3437
                     /label=important segment
     misc_feature    3322..3331
                     /label=SRE(mTOR)
     misc_feature    3343..3352
                     /label=SRE(mTOR)
     misc_feature    3364..3373
                     /label=SRE(mTOR)
     misc_feature    3407..3415
                     /label=E-Box
     misc_feature    3429..3437
                     /label=E-Box
     promoter        3467..3505
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3574..3594
                     /label=181-WKG-593-F
     regulatory      3624..3632
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    3630..4232
                     /label=important segment
     CDS             3630..3698
                     /codon_start=1
                     /label=Secretory (IgK)
                     /label=nonstandard type:  signal
                     /translation="MTSETDTLLLWVLLLWVPGSTGD"
     CDS             3630..3632
                     /codon_start=1
                     /label=Calypso1D6
                     /translation="M"
     CDS             3717..4232
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     primer_bind     4025..4054
                     /label=201-WKG-631-F2
     polyA_signal    4258..4482
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    complement(4566..4594)
                     /label=SleepingBeauty-ITR_Ri
                     /label=nonstandard type:  ITR
     misc_feature    4760..4791
                     /label=SleepingBeauty-ITR-Ro
                     /label=nonstandard type:  ITR
     primer_bind     4853..4872
                     /label=196-WKG-623-F2
     primer_bind     complement(4874..4900)
                     /label=79-WKG-304-F1
     rep_origin      complement(4979..5567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5066..5090)
                     /label=225-WKG-1079-F
     primer_bind     5463..5491
                     /label=197-WKG-623-F3
     CDS             complement(5738..6598)
                     /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"
     primer_bind     6084..6106
                     /label=198-WKG-623-F4
     promoter        complement(join(6599..6628,1..75))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
       61 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gatgcggtgt gaaataccgc
      121 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
      181 attggatccc tatacagttg aagtcggaag tttacataca cttaagttgg agtcattaaa
      241 actcgttttt caactactcc acaaatttct tgttaacaaa caatagtttt ggcaagtcag
      301 ttaggacatc tactttgtgc atgacacaag tcatttttcc aacaattgtt tacagacaga
      361 ttatttcact tataattcac tgtatcacaa ttccagtggg tcagaagttt acatacacta
      421 agttcgactc ctctgcagaa tgcggcgatg tttcggtaag gggtccgcta gaattcgaaa
      481 ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
      541 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta
      601 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
      661 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
      721 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc
      781 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt
      841 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga
      901 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc
      961 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg
     1021 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg
     1081 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg
     1141 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct
     1201 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc
     1261 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc
     1321 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa
     1381 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc
     1441 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg
     1501 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg
     1561 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc
     1621 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aaccggttta
     1681 attaagatat ctgcatctag ggcggccaat tccgcccctc tcccccccgc ccctctccct
     1741 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     1801 atatgtgatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     1861 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     1921 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     1981 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     2041 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     2101 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     2161 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     2221 catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     2281 tcctttgaaa aacacgatga taagcttgcc acaacccaca aggagacgac cttccgtcga
     2341 cgccaccatg ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     2401 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     2461 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     2521 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     2581 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     2641 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     2701 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     2761 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     2821 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     2881 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     2941 cccggtgcct gattcgaagg cctgtcgtgg gggatcaatt cgctcgctga tcagcctcga
     3001 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     3061 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     3121 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     3181 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcctag gagaatctgc
     3241 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtgtattgg
     3301 ccgaaaagtg ccacctgacg tatcacccca cacatctgac gtatcacccc acacatctga
     3361 cgtatcaccc cacacatctg acgtcaggat atcctgcagg acatctatca cgtgactgca
     3421 ggacatctat cacgtgactg caggtcgagc tcggtacccg ggtcgaggta ggcgtgtacg
     3481 gtgggaggcc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     3541 tccacgctgt tttgacctcc atagaagaca ccgggaccga tccagcctcc gcggaattcg
     3601 agctcgcccg gggatcctct agagccacca tgactagtga gacagacaca ctcctgctat
     3661 gggtactgct gctctgggtt ccaggttcca ctggtgacgc tagtggtggt tctggtatgg
     3721 tcttcacact cgaagatttc gttggggact ggcgacagac agccggctac aacctggacc
     3781 aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc
     3841 cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc catgtcatca
     3901 tcccgtatga aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg
     3961 tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg
     4021 acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt
     4081 tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg
     4141 agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg
     4201 gctggcggct gtgcgaacgc attctggcgt gatctagagc tcgctgatca gcctcgactg
     4261 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4321 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     4381 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     4441 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     4501 ccagctgggc ctactagcta ctcgggaccc cttaccgaaa catcgccgca ttctgcagag
     4561 gagtcgagtg tatgtaaact tctgacccac tgggaatgtg atgaaagaaa taaaagctga
     4621 aatgaatcat tctctctact attattctga tatttcacat tcttaaaata aagtggtgat
     4681 cctaactgac ctaagacagg gaatttttac taggattaaa tgtcaggaat tgtgaaaaag
     4741 tgagtttaaa tgtatttggc taaggtgtat gtaaacttcc gacttcaact gtatagggat
     4801 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5401 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5581 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5641 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5701 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5761 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5821 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5881 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5941 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6001 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6061 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6121 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6181 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6241 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6301 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6361 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6421 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6481 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6541 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6601 tcttcctttt tcaatattat tgaagcat
//