LOCUS       sequence-444747-        4743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-2_ST Transposon without spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4743)
  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 4743)
  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-Tc1-2_ST w/o spacer
FEATURES             Location/Qualifiers
     source          1..4743
                     /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        423..922
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(458..481)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     855..874
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      936..945
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             942..1541
                     /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     1441..1461
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1548..1604
                     /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(1605..1607,1608..1610,1611..2324)
                     /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(1605..1607,1608..1610,1611..2321)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1650..1671)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1911..1930)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2258..2279
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2346..2363)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2352..2576
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2846..2862)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2846..2862)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2874..2891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3045..3633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3125..3144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3804..4595,4596..4664))
                     /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     4427..4446
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4665..4743,1..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 tcaattgtac aggtgcatct caataaatta gaatgtcgtg gaaaagttca tttatttcag
      241 taattcaact caaattgtga aactcgtgta ttaaataaat tcaatgcaca cagactgaag
      301 tagtttaagt ctttggttct tttaattgtg atgattttgg ctcacattta acaaaaaccc
      361 accaattcac tatctcaaca aattagaata tggtgacatg ccaatcagct agcaattcta
      421 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct
      481 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag
      541 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt
      601 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc
      661 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct
      721 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag
      781 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc
      841 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc
      901 ttcctcatct ccgggccttt cgacctgcag ccaacgccac catggggacc gagtacaagc
      961 ccacggtgcg cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg
     1021 cgttcgccga ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg
     1081 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg
     1141 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg
     1201 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc
     1261 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg
     1321 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc
     1381 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc
     1441 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg
     1501 aaggaccgcg cacctggtgc atgacccgca agcccggtgc catcgatgcc actaacttct
     1561 ccctgttgaa acaagcaggg gatgtcgaag agaatcccgg gccaatggtg agcaagggcg
     1621 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     1681 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     1741 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     1801 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     1861 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     1921 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     1981 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2041 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2101 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2161 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2221 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2281 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaattaatt aagctcgctg
     2341 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2401 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2461 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2521 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2581 tgaggcgcgc cttattggtc ttatgaagta ttctaatttg ttgagatagt gaattggtgg
     2641 gtttttgtta aatgtgagcc aaaatcatca caattaaaag aaccaaagac ttaaactact
     2701 tcagtctgtg tgcattgaat ttatttaata cacgagtttc acaatttgag ttgaattact
     2761 gaaataaatg aacttttcca cgacattcta atttattgag atgcacctgt agggcccaga
     2821 gacggagtca ctgccaaccg agacggtcat agctgtttcc tgtgtgccgc ttcctcgctc
     2881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2941 gtaatacggt tacccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3781 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3841 atctgtctct ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3901 cgggagggct taccatctgg ccccagtgct gcaataatac cgcgggaccc acgctcaccg
     3961 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4021 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4081 tcgccagtta atagtttgcg caacgttgtt gccatcgcta caggcatcgt ggtatcacgc
     4141 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4201 tcccccatgt tgcgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4261 aagttggccg ccgtgttatc actcatggtt atggcagcac tacataattc tcttactgtc
     4321 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4381 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4441 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4501 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4561 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4621 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4681 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4741 tag
//
