LOCUS       sequence-442033-        6081 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of transposase (CMV 
            promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6081)
  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 6081)
  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-12_AMi
FEATURES             Location/Qualifiers
     source          1..6081
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     121..138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1608..2107
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1643..1666)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2040..2059
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      2121..2130
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2127..2726
                     /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     2626..2646
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2733..2789
                     /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             join(2790..2792,2793..2795,2796..3509)
                     /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             join(2790..2792,2793..2795,2796..3506)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2835..2856)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3096..3115)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3443..3464
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3531..3548)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3537..3761
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4184..4200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4184..4200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4212..4229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4383..4971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4463..4482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5142..5933,5934..6002))
                     /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     5765..5784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6003..6081,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 tcaattgttt ccttgcaggg ctgcccagag gggggggggg gcaactgggg caatttgccc
      241 cgggcccctg caaagcaagg gccccctgcc tcagctcctg tgtgtaccgt tggcccctgc
      301 cctgcgacat cgctcaccag aattcttggt attttgtttt catgaaaaag aggaaattgg
      361 tgtattagag tgtaagatga cactgaatca ttctcaacat gtaattgtga cattatggga
      421 cccgtccccg accatgaagt aaatattata catagttgtg tgtgaaacag atgctaattt
      481 gaggcttaag tgattgttgc attgatgtcc aagggggagt ctaaaccaat cacggtaagg
      541 taacatcttt ctttatctct gggcagcaat agagacaaga ttgatcaggc aggcttatga
      601 ttggcctagc ctagggccta ggccaagtag ccctggttac agagtctgtt gtgaggagag
      661 aaggaacagg gaaggcttta aggagggatc tgtggcagag ccggcctgtt gtcacagaaa
      721 gacctccagg ggaaagagtt caagtcgtga gtgaatgtca gctctgtggg ttcttaaggg
      781 ccaaagtcag aaatgaattt gcaagaatat aataattaaa atgatatata ttgttttctt
      841 tatatttgcc aaggaaaata acaacattag acataacaca tattcaggtt cacatatttc
      901 cccacttcaa aaagtactta aatgcaaagt gtctatataa ctcttgggga aaaatattaa
      961 agtccttctt gattgaagaa gcctgtgctc caaactgcaa atcttgcttt cagtattgca
     1021 gagatgactt gctctcttct ggtccttcaa acagcagcag ttgcttacta aggcccataa
     1081 gaatatgaac agtctcacaa tgcccttaat caaggggtta aaggctaaat atacaagttt
     1141 attctccagc aacagtacaa agacaatcct ccttcctctg tctactctct ctgcagttac
     1201 tttctttttc cttaaacagg ggcagggcct gcttatcctc tgccacaccc acaagcctga
     1261 accaagtaca acccagctta tgggggttac atgagcacca gctaagattt ccaggcaagg
     1321 tcccactgga gagtaatttt ctgtggcagt taccacaggg aaaaggccta ggagtggtgt
     1381 gaggccaggc cttaagggag gaatccttcc ttcctgagta gtggaggagg ccagccagca
     1441 ggccacaacc agaagacctg tgagactgag ccgggcccca ggaaactggt aggagagaga
     1501 agctctgcca gaaggggttt ggaatttgga ggtgctcagg agaagtacaa ccctattcct
     1561 atccccttat ccccgtgtaa gaaaaaaccg gtgctagcaa ttctaccggg taggggaggc
     1621 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
     1681 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
     1741 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
     1801 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
     1861 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
     1921 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
     1981 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
     2041 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
     2101 cctttcgacc tgcagccaac gccaccatgg ggaccgagta caagcccacg gtgcgcctcg
     2161 ccacccgcga cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc
     2221 ccgccacgcg ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag
     2281 aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg
     2341 ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga
     2401 tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag
     2461 gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct
     2521 cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg
     2581 ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct
     2641 acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct
     2701 ggtgcatgac ccgcaagccc ggtgccatcg atgccactaa cttctccctg ttgaaacaag
     2761 caggggatgt cgaagagaat cccgggccaa tggtgagcaa gggcgaggag ctgttcaccg
     2821 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     2881 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     2941 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
     3001 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3061 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3121 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3181 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     3241 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3301 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3361 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     3421 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3481 ctctcggcat ggacgagctg tacaagtaat taattaagct cgctgatcag cctcgactgt
     3541 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3601 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3661 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3721 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cgcgccctcg
     3781 agactttcct cttctcattt tgttcttaag ttcctaactt tgtagcataa gtttaataaa
     3841 tgtttcactt gcaaactgtt aatgtcattt tgctttagta acctgtctcc aaggtaaatt
     3901 ctctctgcct agccctcctt gccacgggaa caagcgggga aagcccactt gttcccactt
     3961 cccagggcaa gggaaggagg agccgcatgt gagaattctc acatgcgatg cgtgcgtgca
     4021 agaattctcg cacgtggtcc cccagctttg tacacacaca cacgggtcct tgtagggggc
     4081 agtggtgggg cccccaacat cacacctgcc ccaggcccca taaaccctct gggcagccct
     4141 gtttccttgg ggcccagaga cggagtcact gccaaccgag acggtcatag ctgtttcctg
     4201 tgtgccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4261 tcagctcact caaaggcggt aatacggtta cccacagaat caggggataa cgcaggaaag
     4321 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4381 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4441 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4501 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4561 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4621 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4681 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4741 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4801 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4861 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4921 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4981 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5041 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5101 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5161 gaggcaccta tctcagcgat ctgtctcttt cgttcatcca tagttgcctg actccccgtc
     5221 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aataataccg
     5281 cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5341 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5401 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca
     5461 ggcatcgtgg tatcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5521 tcaaggcgag ttacatgatc ccccatgttg cgcaaaaaag cggttagctc cttcggtcct
     5581 ccgatcgttg tcagaagtaa gttggccgcc gtgttatcac tcatggttat ggcagcacta
     5641 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5701 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5761 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5821 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5881 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5941 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6001 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6061 tacatatttg aatgtattta g
//
