LOCUS       sequence-442204-        6029 bp    DNA     circular SYN 15-APR-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 6029)
  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 6029)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-13_AMi
FEATURES             Location/Qualifiers
     source          1..6029
                     /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        1471..1970
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1506..1529)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1903..1922
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1984..1993
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1990..2589
                     /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     2489..2509
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2596..2652
                     /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(2653..2655,2656..2658,2659..3372)
                     /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(2653..2655,2656..2658,2659..3369)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2698..2719)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2959..2978)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3306..3327
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3394..3411)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3400..3624
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4132..4148)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4132..4148)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4160..4177)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4331..4919)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4411..4430)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5090..5881,5882..5950))
                     /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     5713..5732
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5951..6029,1..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 tcaattggtc ataagcagag atgtgggttt tcgcggaaaa ttttccagtg ctttattttt
      241 ccatgggaaa ttttccattt ctaaaaattc attgtttcat aaaattcatt agtaacaatg
      301 aatggatgcc aatacaatat taggcaagct tggcagtttc aaagttgtaa ttaatgtttt
      361 tcaggtgatt atccctagga agtgtgtgag agagtacata tatgttttat tgatttgtaa
      421 acaggagggt aaatacactc acataataat ccaaaccaaa aatcaagtta tgaattcacc
      481 taattggctg tgcaggtcaa atcaaaacca aagctaaaat tcttatttca tctcagactg
      541 caagtgaaag caaaaccaaa actaaaactg tgattaattt ttgtttcact ttcagttcag
      601 caaacacaca gagcagtgtc tcatttcttt tcttgagttg gggaagttgg aagtgtttgc
      661 ataagatctg ttagtttttt agtgcttttg gtttgaagta gtcataataa tgcctaatca
      721 gaaaaaaata gtgtgtggca tcattatagt gaagttcaag ccactgaagg aaggaagaaa
      781 gttggtagat gtaaatattg cgggcaggag tatgctaaca acgcaacaag aatgaagaaa
      841 catttaataa catgtagaaa ctgtcctgaa agagtacaag ccatatacag aaagctaata
      901 caacaaagtg gtggaacaac atcatctgag aaaagtctac ttcaggaatc ctatgtaagt
      961 aaacaaatta atatgttaaa caacgcacag tatagactag catctatatt gttttaaaac
     1021 attataattt taaaagttgt taatgcaact ctggactatt atttatttat ttatttattt
     1081 aaaatatttg tatcctgctt ttctgcttgc actaagcacc caaagtggct taggcttaac
     1141 aatatatttt ttaaattaca taatagataa atagttaact atttcaaccc agcagagtga
     1201 gcttagtcct ctggccgatg gcagaggcca gactatactt ccctttggct ctgcagtccc
     1261 agggcaactg agcagcaagt tgagcagcag agtgttcttt ctggcaggga ggggggaagc
     1321 aagctccctg ccgcttctcc ttctccttct gatggatgag gccagactag aataggctaa
     1381 aataaaaatc ttttccccct tttgttgcag gaacttggta gctcaagtga aatagcagat
     1441 ttgcaaaaaa ccggtgctag caattctacc gggtagggga ggcgcttttc ccaaggcagt
     1501 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     1561 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     1621 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     1681 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     1741 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     1801 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     1861 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     1921 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     1981 aacgccacca tggggaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     2041 ccccgggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     2101 gtcgacccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     2161 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     2221 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     2281 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     2341 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     2401 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     2461 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     2521 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     2581 cccggtgcca tcgatgccac taacttctcc ctgttgaaac aagcagggga tgtcgaagag
     2641 aatcccgggc caatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2701 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2761 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2821 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2881 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2941 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3001 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3061 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3121 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3181 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3241 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3301 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3361 ctgtacaagt aattaattaa gctcgctgat cagcctcgac tgtgccttct agttgccagc
     3421 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3481 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3541 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3601 ctggggatgc ggtgggctct atggcttctg aggcgcgccc tcgagactct gttgaagtag
     3661 atgagtggac aaccacggca tatcatgcac agttctttgg gtgtgctaaa attagtgtgt
     3721 caacagtttt cagtgccttc gtttttttcc ttaaatttaa acaaacaatg taagggaagt
     3781 acatgctatt gtgtattgtc tttacatttt tacaagtatt ccaataaaaa taatttcttt
     3841 ccacataaag ctttgaattt gtttgaatat aacatttaaa ccacattaag tgtgctgtat
     3901 ttaatttttt tcctaatcaa atatttcagt tcaaatactt gtggctattg ggacataaaa
     3961 ttaacatggg ggtacttttt tagttttgtt tactaaatat aagtcaaata aacatgctaa
     4021 atcagatcac gctctattta gggcattgga aaatttccat caaaattttc cggaaaaccc
     4081 acatctctgg tcataagggg cccagagacg gagtcactgc caaccgagac ggtcatagct
     4141 gtttcctgtg tgccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4201 gagcggtatc agctcactca aaggcggtaa tacggttacc cacagaatca ggggataacg
     4261 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4321 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4381 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4441 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4501 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4561 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4621 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4681 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4741 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4801 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4861 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4921 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4981 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5041 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5101 taatcagtga ggcacctatc tcagcgatct gtctctttcg ttcatccata gttgcctgac
     5161 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5221 taataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5281 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5341 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5401 tcgctacagg catcgtggta tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5461 cccaacgatc aaggcgagtt acatgatccc ccatgttgcg caaaaaagcg gttagctcct
     5521 tcggtcctcc gatcgttgtc agaagtaagt tggccgccgt gttatcactc atggttatgg
     5581 cagcactaca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5641 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5701 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5761 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5821 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5881 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5941 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6001 tgagcggata catatttgaa tgtatttag
//
