LOCUS       sequence-443383-        4768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-1_Xt Transposon without spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4768)
  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 4768)
  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 w/o spacer
FEATURES             Location/Qualifiers
     source          1..4768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     231..250
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     490..511
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(557..613)
                     /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             complement(620..1219)
                     /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     complement(700..720)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     regulatory      1216..1225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1239..1738)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1287..1306)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1680..1703
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(2022..2039)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2022..2038)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2048..2066)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      2101..2130
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      2152..2739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2640..2659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2751..2778)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(2802..2993)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(2994..3785,3786..3854))
                     /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     3617..3636
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3855..3957)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3958..3989)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     4041..4058
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4070..4086
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4070..4086
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    4353..4577
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     4566..4583
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(4605..4768,1..550,551..553,554..556))
                     /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             complement(join(4608..4768,1..550,551..553,554..556))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4650..4671)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg cggatcttga
       61 agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg tggctgttgt
      121 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
      181 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
      241 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
      301 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct gaagcactgc acgccgtagg
      361 tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatgaact
      421 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
      481 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
      541 cgcccttgct caccattggc ccgggattct cttcgacatc ccctgcttgt ttcaacaggg
      601 agaagttagt ggcatcgatg gcaccgggct tgcgggtcat gcaccaggtg cgcggtcctt
      661 cgggcacctc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag gggaggttgc
      721 ggggcgcgga ggtctccagg aaggcgggca ccccggcgcg ctcggccgcc tccactccgg
      781 ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg ccgacggtgg
      841 ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc atctgttgct
      901 gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg gcgaacaccg
      961 cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg tcgtccgcga
     1021 cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc agctcggtga
     1081 cccgctcgat gtggcggtcc gggtcgacgg tgtggcgcgt ggcggggtag tcggcgaacg
     1141 cggcggcgag ggtgcgtacg gcccggggga cgtcgtcgcg ggtggcgagg cgcaccgtgg
     1201 gcttgtactc ggtccccatg gtggcgttgg ctgcaggtcg aaaggcccgg agatgaggaa
     1261 gaggagaaca gcgcggcaga cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag
     1321 gaccttcggg cgcccgcccc gcccctgagc ccgcccctga gcccgccccc ggacccaccc
     1381 cttcccagcc tctgagccca gaaagcgaag gagcaaagct gctattggcc gctgccccaa
     1441 aggcctaccc gcttccattg ctcagcggtg ctgtccatct gcacgagact agtgagacgt
     1501 gctacttcca tttgtcacgt cctgcacgac gcgagctgcg gggcgggggg gaacttcctg
     1561 actaggggag gagtagaagg tggcgcgaag gggccaccaa agaacggagc cggttggcgc
     1621 ctaccggtgg atgtggaatg tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc
     1681 agcggggctg ctaaagcgca tgctccagac tgccttggga aaagcgcctc ccctacccgg
     1741 tagaattgct agcagggggg caatcacttt ttcacacagg gccatgtaga tttggagttt
     1801 tttttctccc ttaataacgt aaaccttcat ttaaaaactg cattttgtgt tcaattatgt
     1861 tatctttgac taatagttaa cggtttttga tgagcagaaa catttaagtg tgacaaacat
     1921 gcaaaagaat aagaaatcag gaagggggca aatagttttt cacaccactg taaagcttag
     1981 agacgcaaga cactgcggat cgagacgcat tgcgtggttc gactggccgt cgttttacaa
     2041 cgaaagggcc tcgtgatacg cctattgact ccagcgtaac tggactgcaa tcaactcact
     2101 ggctcacctt cacgggtggg cctttcttcg gtagaaaatc aaaggatctt cttgagatcc
     2161 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2221 ttgtttgccg gatcaagagc taccaactct ttttccgagg taactggctt cagcagagcg
     2281 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2341 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2401 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2461 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2521 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2581 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2641 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcatcga
     2701 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcagaaagg
     2761 cccacccgaa ggtgagccag gtgattacat ttgggccctc atcagaggtt ttcaccgtca
     2821 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     2881 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     2941 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cacttaccaa
     3001 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3061 tgactccccg tcgtgtagat aactacgatg cgggagggct taccatctgg ccccagttct
     3121 gcaatgatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3181 gccgggaggg ccgagcgcag aagtgatcct gcaactttat ccgcctccat ccagtctatt
     3241 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3301 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     3361 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     3421 tccttcggtc ctccgatcgt tgccagaagt aagttggccg cagtgttatc actcatggtt
     3481 atggcagcac tgcataattc tcttactgtc atgccatccg tgagatgctt ttctgtgact
     3541 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     3601 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     3661 ggaaaacgtt cttcggggcg taaactctca aggatcttac cgctgttgag atccagttcg
     3721 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     3781 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     3841 tgttgaatac tcattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg
     3901 cccggtagtg atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaagt
     3961 caaaagcctc cggtcggagg cttttgactt tctgctatgg aggtcaggta tgatttaaat
     4021 ggtcagtaac gaccgagcgc agcgagtcag tgagcgagga agcggcacac aggaaacagc
     4081 tatgaccgtc tcggttggca gtgactccgt ctctgggccc tacagtggtg tgaaaaacta
     4141 tttgccccct tcctgatttc ttattctttt gcatgtttgt cacacaaaat gtttctgatc
     4201 atcaaacaca tttaactatt agtcaaagat aacacaagta aacacaaaat gcagttttta
     4261 aatgagggtt tttattattt agggagaaaa aaaatccaaa cctacatggc cctgtgtgaa
     4321 aaagtaattg ccccctgggc gcgcctcaga agccatagag cccaccgcat ccccagcatg
     4381 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     4441 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     4501 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     4561 gcaactagaa ggcacagtcg aggctgatca gcgagcttaa ttaattactt gtacagctcg
     4621 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     4681 ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc
     4741 agcacggggc cgtcgccgat gggggtgt
//
