LOCUS       sequence-444759-        4937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-1_Xt Transposon with copGFP mammalian 
            expression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4937)
  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 4937)
  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: pQLL-Tc1-1_Xt[EF1-alpha-copGFP]
FEATURES             Location/Qualifiers
     source          1..4937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     120..139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(231..258)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(282..473)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(474..1265,1266..1334))
                     /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     1097..1116
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1335..1437)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(1438..1469)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     1521..1538
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1550..1566
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1550..1566
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1953..2034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1990..2009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2044..2063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(2077..2742)
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             complement(2077..2742)
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     promoter        complement(2784..3962)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2793..3731
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(2816..2836)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(4439..4456)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4439..4455)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4465..4483)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      4518..4547
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      join(4569..4937,1..219)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
       61 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
      121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga
      181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg
      241 cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca
      301 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
      361 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
      421 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa
      481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
      541 tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagttct
      601 gcaatgatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
      661 gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt
      721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
      781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
      841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
      901 tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt
      961 atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact
     1021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     1081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     1141 ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg
     1201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     1261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     1321 tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     1381 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt
     1441 caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat
     1501 ggtcagtaac gaccgagcgc agcgagtcag tgagcgagga agcggcacac aggaaacagc
     1561 tatgaccgtc tcggttggca gtgactccgt ctctgggccc tacagtggtg tgaaaaacta
     1621 tttgccccct tcctgatttc ttattctttt gcatgtttgt cacacaaaat gtttctgatc
     1681 atcaaacaca tttaactatt agtcaaagat aacacaagta aacacaaaat gcagttttta
     1741 aatgagggtt tttattattt agggagaaaa aaaatccaaa cctacatggc cctgtgtgaa
     1801 aaagtaattg ccccctgaac ctcgagggcg cgccctcgcc ctttggtctc cccctcttaa
     1861 gtaccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     1921 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     1981 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2041 gttgtggttt gtccaaactc atcgagctcg agatctggcg aaggcgatgg gggtcttgaa
     2101 ggcgtgctgg tactccacga tgcccagctc ggtgttgctg tgcagctcct ccacgcggcg
     2161 gaaggcgaac atggggcccc cgttctgcag gatgctgggg tggatggcgc tcttgaagtg
     2221 catgtggctg tccaccacga agctgtagta gccgccgtcg cgcaggctga aggtgcgggc
     2281 gaagctgccc accagcacgt tatcgcccat ggggtgcagg tgctccacgg tggcgttgct
     2341 gcggatgatc ttgtcggtga agatcacgct gtcctcgggg aagccggtgc ccaccacctt
     2401 gaagtcgccg atcacgcggc cggcctcgta gcggtagctg aagctcacgt gcagcacgcc
     2461 gccgtcctca tacttctcga tgcgggtgtt ggtgtagccg ccgttgttga tggcgtgcag
     2521 gaaggggttc tcgtagccgc tggggtaggt gccgaagtgg tagaagccgt agcccatcac
     2581 gtggctcagc aggtaggggc tgaaggtcag ggcgcctttg gtgctcttca tcttgttggt
     2641 catgcggccc tgctcggggg tgccctctcc gccgcccacc agctcgaact ccacgccgtt
     2701 cagggtgccg gtgatgcggc actcgatctt catggcgggc atggtggcga ccggtagcgc
     2761 tagcggcttc ggtaccggat ccctcacgac acctgaaatg gaagaaaaaa actttgaacc
     2821 actgtctgag gcttgagaat gaaccaagat ccaaactcaa aaagggcaaa ttccaaggag
     2881 aattacatca agtgccaagc tggcctaact tcagtctcca cccactcagt gtggggaaac
     2941 tccatcgcat aaaacccctc cccccaacct aaagacgacg tactccaaaa gctcgagaac
     3001 taatcgaggt gcctggacgg cgcccggtac tcagtggagt cacatgaagc gacggctgag
     3061 gacggaaagg cccttttcct ttgtgtgggt gactcacccg cccgctctcc cgagcgccgc
     3121 gtcctccatt ttgagctccc tgcagcaggg ccgggaagcg gccatctttc cgctcacgca
     3181 actggtgccg accgggccag ccttgccgcc cagggcgggg cgatacacgg cggcgcgagg
     3241 ccaggcacca gagcaggccg gcgagcttga gactaccccc gtccgattct cggtggccgc
     3301 gctcgcaggc cccgcctcgc cgaacatgtg cgctgggacg cacgggcccc gtcgccgccc
     3361 gcggccccaa aaaccgaaat accagtgtgc agatcttggc ccgcatttac aagactatct
     3421 tgccagaaaa aaagcgtcgc agcaggtcat caaaaatttt aaatggctag agacttatcg
     3481 aaagcagcga gacaggcgcg aaggtgccac cagattcgca cgcggcggcc ccagcgccca
     3541 ggccaggcct caactcaagc acgaggcgaa ggggctcctt aagcgcaagg cctcgaactc
     3601 tcccacccac ttccaacccg aagctcggga tcaagaatca cgtactgcag ccaggtggaa
     3661 gtaattcaag gcacgcaagg gccataaccc gtaaagaggc caggcccgcg ggaaccacac
     3721 acggcactta cctgtgttct ggcggcaaac ccgttgcgaa aaagaacgtt cacggcgact
     3781 actgcactta tatacggttc tcccccaccc tcgggaaaaa ggcggagcca gtacacgaca
     3841 tcactttccc agtttacccc gcgccacctt ctctaggcac cgggtcaatt gccgacccct
     3901 ccccccaact tctcggggac tgtgggcgat gtgcgctctg cccactgacg ggcaccggag
     3961 ccactagtca ataatcaatg cctctagcac cggtgatgtc tagcttttga ggtgcttttg
     4021 gtctacttct ctgtgtcagg tagctcctat ttaagtgatt tcttgattga aacaggtgtg
     4081 gcagtaatca ggcctggggg tgactacaga aattgatatt gaaattgaaa attgaaatgt
     4141 gataaaccac agttaagtta ttttttaaca aggggggcaa tcactttttc acacagggcc
     4201 atgtagattt ggagtttttt ttctccctta ataacgtaaa ccttcattta aaaactgcat
     4261 tttgtgttca attatgttat ctttgactaa tagttaacgg tttttgatga gcagaaacat
     4321 ttaagtgtga caaacatgca aaagaataag aaatcaggaa gggggcaaat agtttttcac
     4381 accactgtaa agcttagaga cgcaagacac tgcggatcga gacgcattgc gtggttcgac
     4441 tggccgtcgt tttacaacga aagggcctcg tgatacgcct attgactcca gcgtaactgg
     4501 actgcaatca actcactggc tcaccttcac gggtgggcct ttcttcggta gaaaatcaaa
     4561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaggtaa
     4681 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4741 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4801 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4861 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4921 gaacgaccta caccgaa
//
