LOCUS       sequence-445649-        8085 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-7_PM Transposon with 5 kb cargo gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8085)
  AUTHORS   Zhang T, Tan S, Tang N, Li Y, Zhang C, Sun J, Guo Y, Gao H, Cai Y, 
            Sun W, Wang C, Fu L, Ma H, Wu Y, Hu X, Zhang X, Gee P, Yan W, Zhao 
            Y, Chen Q, Guo B, Wang H, Zhang YE
  TITLE     Heterologous survey of 130 DNA transposons in human cells highlights
            their functional divergence and expands the genome engineering 
            toolbox.
  JOURNAL   Cell. 2024 Jul 11;187(14):3741-3760.e30. doi: 
            10.1016/j.cell.2024.05.007. Epub 2024 Jun 5.
  PUBMED    38843831
REFERENCE   2  (bases 1 to 8085)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pMV-hAT-7_PM-5 kb
FEATURES             Location/Qualifiers
     source          1..8085
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    51..132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(88..107)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1675..1698)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      1698..1707
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1722..2900)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1731..2669
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(1754..1774)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     polyA_signal    2946..3170
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3159..3176
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3198..3911,3912..3914,3915..3917))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3201..3911,3912..3914,3915..3917))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3243..3264)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3592..3611
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3851..3872
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(3918..3974)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(3981..4580)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4061..4081)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     regulatory      4577..4586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4600..5095)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4648..4667)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     5041..5064
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     regulatory      5111..5120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5515..5532)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5515..5531)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5541..5559)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5627..5731
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5732..5800,5801..6592)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5950..5969)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6763..7351
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7252..7271
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7505..7522
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7534..7550
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7534..7550
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
       61 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
      121 ttttttcact gcattctagt tgtggtttgt ccaaactcaa tcgatgcgag ggggcagggc
      181 ctgcatgtga agggcgtcgt aggtgtcctt ggtggctgta ctgagaccct ggtaaaggcc
      241 atcgtgcccc ttgcccctcc ggcgctcgcc tttcatccca atctcactgt aggcctccgc
      301 catcttatct ttctgcagtt cattgtacag gccttcctga gggttcttcc ttctcggctt
      361 tccccccatc tcagggtccc ggccacgtct cttgtccaaa acatcgtact cctctcttcg
      421 tcctagattg agctcgttat agagctggtt ctggccctgc tggtacgcgg gggcgtctgc
      481 gctcctgctg aacttcactc tggagcgata ggctgcgaag tcgcgtggtg gggcataggg
      541 ctggtaatgc ttgcgggtgg gcccggggcg gcggggagtc atgttcatgt agtcactgtg
      601 caggagcctg ctcctcttac tcctcaccca gaaaataata aaggccactg ttactagcaa
      661 gctatagcaa gccaggactc ccccaaccac caccagcacc caaaaatcac aggcgaagtc
      721 cagccccctc gtgtgcactg cgccccccgc cgctggccgg cacgcctctg ggcgcaggga
      781 caggggctgc gacgcgatgg tgggcgccgg tgttggtggt cgcggcgctg gcgtcgtggt
      841 cagcactgtc agttgcgtgc ctccaccgaa aacagcggct gaactatgcc aaatcataca
      901 gtaataatcg gcctcgtcct cggacctgag acctgagatg agcagcacac ccgcattggc
      961 gcttgcgtct ttagatccac taaaccttga tggaactcca gatccctgct gcttatctga
     1021 gtcgctcttg taattgagaa ggtattgggg aggagaacct ggtttttgct gataccagta
     1081 gatcctgtaa ggccctacat tgattcctga cctcagtgta cacgtcaaag aagcggaggc
     1141 accggggctt gcactgaggg aggatgattg cgtgagaaca ggttggctac ccccccctcc
     1201 ggacccgccc ccaccggaac ctccaccgcc tgagcccagg atcccactgg acacggtgac
     1261 ggttgtacct tgtccccaaa cgtccatacc atagtagtaa gtcatcatac cccgcgcaca
     1321 atagtaaaca gctgtgtctt cgggagttac actattaagc tgaagagaga actggttttt
     1381 ggaagtatct ggatttatag acatcctgct tttcacactt acagcataat cattatacca
     1441 tttactgcgg tagtatgttc taccgagcca ttccaaccct cgactggggg actgcctgat
     1501 ccaattccat gtcgcggagt tactactcac tgaatcacct gatatggcac aggtaagact
     1561 aagagtctgg gaaggagtga ccagaccggg gccagattgt tggagttgca cttgcggcct
     1621 ggcggcgtgg agcagcaagg ccagcggcag gagcaaggcg gtcactggta aggccttatc
     1681 gtcatcgtct ttgtaatcca tggtggcgac cggtggatcc ctcacgacac ctgaaatgga
     1741 agaaaaaaac tttgaaccac tgtctgaggc ttgagaatga accaagatcc aaactcaaaa
     1801 agggcaaatt ccaaggagaa ttacatcaag tgccaagctg gcctaacttc agtctccacc
     1861 cactcagtgt ggggaaactc catcgcataa aacccctccc cccaacctaa agacgacgta
     1921 ctccaaaagc tcgagaacta atcgaggtgc ctggacggcg cccggtactc agtggagtca
     1981 catgaagcga cggctgagga cggaaaggcc cttttccttt gtgtgggtga ctcacccgcc
     2041 cgctctcccg agcgccgcgt cctccatttt gagctccctg cagcagggcc gggaagcggc
     2101 catctttccg ctcacgcaac tggtgccgac cgggccagcc ttgccgccca gggcggggcg
     2161 atacacggcg gcgcgaggcc aggcaccaga gcaggccggc gagcttgaga ctacccccgt
     2221 ccgattctcg gtggccgcgc tcgcaggccc cgcctcgccg aacatgtgcg ctgggacgca
     2281 cgggccccgt cgccgcccgc ggccccaaaa accgaaatac cagtgtgcag atcttggccc
     2341 gcatttacaa gactatcttg ccagaaaaaa agcgtcgcag caggtcatca aaaattttaa
     2401 atggctagag acttatcgaa agcagcgaga caggcgcgaa ggtgccacca gattcgcacg
     2461 cggcggcccc agcgcccagg ccaggcctca actcaagcac gaggcgaagg ggctccttaa
     2521 gcgcaaggcc tcgaactctc ccacccactt ccaacccgaa gctcgggatc aagaatcacg
     2581 tactgcagcc aggtggaagt aattcaaggc acgcaagggc cataacccgt aaagaggcca
     2641 ggcccgcggg aaccacacac ggcacttacc tgtgttctgg cggcaaaccc gttgcgaaaa
     2701 agaacgttca cggcgactac tgcacttata tacggttctc ccccaccctc gggaaaaagg
     2761 cggagccagt acacgacatc actttcccag tttaccccgc gccaccttct ctaggcaccg
     2821 ggtcaattgc cgacccctcc ccccaacttc tcggggactg tgggcgatgt gcgctctgcc
     2881 cactgacggg caccggagcc tcacgttaat taaccaaggt acccattggc ggccgccctc
     2941 agaagccata gagcccaccg catccccagc atgcctgcta ttgtcttccc aatcctcccc
     3001 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     3061 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     3121 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga
     3181 tcagcgagct taattaatta cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     3241 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     3301 gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     3361 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg
     3421 aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg
     3481 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     3541 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     3601 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     3661 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag
     3721 gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     3781 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     3841 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     3901 tcgcccttgc tcaccattgg cccgggattc tcttcgacat cccctgcttg tttcaacagg
     3961 gagaagttag tggcatcgat ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct
     4021 tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg
     4081 cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg
     4141 gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac gccgacggtg
     4201 gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc
     4261 tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc
     4321 gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg
     4381 acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg
     4441 acccgctcga tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta gtcggcgaac
     4501 gcggcggcga gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag gcgcaccgtg
     4561 ggcttgtact cggtccccat ggtggcgttg gctgcaggtc gaaaggcccg gagatgagga
     4621 agaggagaac agcgcggcag acgtgcgctt ttgaagcgtg cagaatgccg ggcctccgga
     4681 ggaccttcgg gcgcccgccc cgcccctgag cccgcccctg agcccgcccc cggacccacc
     4741 ccttcccagc ctctgagccc agaaagcgaa ggagcaaagc tgctattggc cgctgcccca
     4801 aaggcctacc cgcttccatt gctcagcggt gctgtccatc tgcacgagac tagtgagacg
     4861 tgctacttcc atttgtcacg tcctgcacga cgcgagctgc ggggcggggg ggaacttcct
     4921 gactagggga ggagtagaag gtggcgcgaa ggggccacca aagaacggag ccggttggcg
     4981 cctaccggtg gatgtggaat gtgtgcgagg ccagaggcca cttgtgtagc gccaagtgcc
     5041 cagcggggct gctaaagcgc atgctccaga ctgccttggg aaaagcgccc tacccggtag
     5101 aattgtatac ccatggcggc cgggagcatg cgacgtcggg gcccgctagc accggtttta
     5161 cagaagggct cacagggcaa agcgaaaaag tgaagggagc accgtatcta actcagcaat
     5221 gtaaacaaac cctgaaaatg gcgctgacgg cccacaatgc attgcgagat catgtgccct
     5281 ttcgagccct atacaccaca ctcccggtca aagtgcgcca tctaatgggg aaatgggggc
     5341 attacaggag aaaggtaaaa tttaatttaa taataatatt tggacaagtt cggcgggccg
     5401 gattaaaaag cccaacgggc cgtatacggc ccgggggccg tactttgccc atgtctggtc
     5461 tagacaagct tagagacgca agacactgcg gatcgagacg cattgcgtgg ttcgactggc
     5521 cgtcgtttta caacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     5581 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5641 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5701 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5761 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5821 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5881 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5941 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6001 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6061 tcttacggat ggcatgacag taagagaatt atgtagtgct gccataacca tgagtgataa
     6121 cacggcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6181 gcgcaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6241 cataccaaac gacgagcgtg ataccacgat gcctgtagcg atggcaacaa cgttgcgcaa
     6301 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6361 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6421 tgataaatct ggagccggtg agcgtgggtc ccgcggtatt attgcagcac tggggccaga
     6481 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6541 acgaaagaga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6601 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6661 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6721 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6781 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6841 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6901 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6961 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7021 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7081 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7141 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7201 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7261 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7321 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7381 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7441 gggtaaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     7501 gcgcagcgag tcagtgagcg aggaagcggc acacaggaaa cagctatgac cgtctcggtt
     7561 ggcagtgact ccgtctctgg gcccgtctag accaggggtg ggcaaatttt ttgactcgcg
     7621 ggccacaatg ggttctaaaa tttgacagag gggccgggcc aggaacagat ggatggagtg
     7681 tttgtgtgaa ctaatataaa tgacatgtaa aaatcattac atgaaaggat ttggccttta
     7741 acaggtagca aagcatttat ttgcaaggca atttaacaaa ttgcacgaac atgaacaata
     7801 caaaaacatg tcgtaacgtg actgatcaaa catcatgaaa acatctgagg cctgcgccca
     7861 aaccaatgac ctcacttgaa cttctttatt atctacatta acaggttaga gcagccacgc
     7921 aggctgtgtg tcggagattt ggctccaacc tcgctgcagc aacacatcat cactttcagt
     7981 tctcgagggc gcgccactag tgaattctgc aggtcgacca tatgtgtaca agcgcttcag
     8041 aaggtaccac atttgtagag gttttacttg ctttaaaaaa cctcc
//
