LOCUS       sequence-442074-        4975 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-8_Xt Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4975)
  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 4975)
  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-Tc1-8_Xt
FEATURES             Location/Qualifiers
     source          1..4975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..82
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     91..108
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     92..108
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        580..1079
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(615..638)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1012..1031
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1093..1102
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1099..1698
                     /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     1598..1618
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1705..1761
                     /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(1762..1764,1765..1767,1768..2481)
                     /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(1762..1764,1765..1767,1768..2478)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1807..1828)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2068..2087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2415..2436
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2503..2520)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2509..2733
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3019..3035)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3019..3035)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3047..3064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3116..3147
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3148..3250
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(3251..3319,3320..4111)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAELGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3469..3488)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4112..4303
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      4327..4354
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(4366..4953)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4446..4465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(join(4975,1..29))
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
       61 gtcaataggc gtatcacgag gccctttcgt tgtaaaacga cggccagtcg aaccacgcaa
      121 tgcgtctcga tccgcagtgt cttgcgtctc tctgcagtac agtcatatga aaaagtttgg
      181 gaacccctct cagcctgcat aataatttac tccactttca acaaaaaaag ataacagtgg
      241 tatgtctttc atttcccagg aacatctgag tactggggtg ttttctgaac aaagattttt
      301 agtgaagcag tatttagttg tatgaaatta aatcaaatgt gaaaaactgg ctgtgcaaaa
      361 atttgggtac ccttgtaatt ttgctaattt gaatgcatgt aactgctcaa tactgattac
      421 tggcaacacc aaattggttg gattagcttg ttaagccttg aacttcatag gcaggtgtgt
      481 ccaatcatga gaaaaggtat ttaaggtggc caattgcaag ttgtgcttct gtttgactct
      541 cctctgaaga gtgacagcac cggtgctagc aattctaccg ggtaggggag gcgcttttcc
      601 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
      661 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
      721 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
      781 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
      841 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
      901 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
      961 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
     1021 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
     1081 cctgcagcca acgccaccat ggggaccgag tacaagccca cggtgcgcct cgccacccgc
     1141 gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     1201 cgccacaccg tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     1261 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     1321 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     1381 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     1441 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     1501 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     1561 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     1621 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     1681 acccgcaagc ccggtgccat cgatgccact aacttctccc tgttgaaaca agcaggggat
     1741 gtcgaagaga atcccgggcc aatggtgagc aagggcgagg agctgttcac cggggtggtg
     1801 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1861 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1921 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1981 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2041 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2101 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2161 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2221 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2281 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2341 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2401 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2461 atggacgagc tgtacaagta attaattaag ctcgctgatc agcctcgact gtgccttcta
     2521 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2581 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     2641 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     2701 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcgcgccct cgagtgtaat
     2761 atctctgttg gggtgcccaa atttatgcac ctgtctaatt ttgttatgat gcatattgca
     2821 tattttctgt taatccaata aactttatgt cactgctgaa atactactgt ttccataagg
     2881 catgttatat attaaaagga agttgctact ttgaaagctc agccaatgat aaacaaaact
     2941 ccaaagaatt aagaggggtt cccaaacttt ttcatatgac tgtagggccc agagacggag
     3001 tcactgccaa ccgagacggt catagctgtt tcctgtgtgc cgcttcctcg ctcactgact
     3061 cgctgcgctc ggtcgttact gaccatttaa atcatacctg acctccatag cagaaagtca
     3121 aaagcctccg accggaggct tttgacttga tcggcacgta agaggttcca actttcacca
     3181 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     3241 ggaagctaaa atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3301 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3361 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3421 tttacgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3481 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3541 gaatgacttg gttgagtact caccagtcac agaaaagcat ctcacggatg gcatgacagt
     3601 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3661 ggcaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3721 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3781 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3841 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggatc
     3901 acttctgcgc tcggccctcc cggctggctg gtttattgct gataaatctg gagccggtga
     3961 gcgtgggtcc cgcggtatca ttgcagaact ggggccagat ggtaagccct cccgcatcgt
     4021 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4081 gataggtgcc tcactgatta agcattggta agtgaccaaa caggaaaaaa ccgcccttaa
     4141 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     4201 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     4261 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgatgagggc ccaaatgtaa
     4321 tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc ataggctccg
     4381 cccccctgac gagcatcaca aaaatcgatg ctcaagtcag aggtggcgaa acccgacagg
     4441 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4501 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4561 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4621 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4681 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4741 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4801 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttacctcgg aaaaagagtt
     4861 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4921 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatttt ctacc
//
