LOCUS       sequence-443393-        4749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-8B_DR Transposon without spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4749)
  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 4749)
  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-Tc1-8B_DR w/o spacer
FEATURES             Location/Qualifiers
     source          1..4749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(124..226)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(227..258)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     310..327
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     339..355
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     339..355
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    620..844
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     833..850
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(872..1585,1586..1588,1589..1591))
                     /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(875..1585,1586..1588,1589..1591))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(917..938)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1266..1285
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1525..1546
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(1592..1648)
                     /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(1655..2254)
                     /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(1735..1755)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     regulatory      2251..2260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2274..2773)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2322..2341)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2715..2738
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     complement(3040..3057)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3040..3056)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3066..3084)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      3119..3148
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      3170..3757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3658..3677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3769..3796)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(3820..4011)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(4012..4749,1..54,55..123))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4635..4654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
       61 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      121 cattttagct tccttagctc ctgaaaatct cgataactca aaaaatacgc ccggtagtga
      181 tcttatttca ttatggtgaa agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc
      241 ggtcggaggc ttttgacttt ctgctatgga ggtcaggtat gatttaaatg gtcagtaacg
      301 accgagcgca gcgagtcagt gagcgaggaa gcggcacaca ggaaacagct atgaccgtct
      361 cggttggcag tgactccgtc tctgggccct acagcgggga aaataagtat ttgacacatc
      421 agcattttta tcagtaaggg gatttctaag tgggctactg acacaaaatt cctaccagat
      481 gtagccatca agccaaatat tgaattcata caaagaaatc agaacattta agtatacaag
      541 ttgagtcata ataaataaag tgaaatgaca cagggaataa gtattgaaca cactttacct
      601 cgagggcgcg cctcagaagc catagagccc accgcatccc cagcatgcct gctattgtct
      661 tcccaatcct cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta
      721 ctcagacaat gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct
      781 tccagggtca aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc
      841 acagtcgagg ctgatcagcg agcttaatta attacttgta cagctcgtcc atgccgagag
      901 tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc tcgttggggt
      961 ctttgctcag ggcggactgg gtgctcaggt agtggttgtc gggcagcagc acggggccgt
     1021 cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg tcctcgatgt
     1081 tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc atgatataga
     1141 cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg tcctccttga
     1201 agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac ttcacctcgg
     1261 cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg acgtagcctt
     1321 cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag cggctgaagc
     1381 actgcacgcc gtaggtcagg gtggtcacga gggtgggcca gggcacgggc agcttgccgg
     1441 tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg
     1501 acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc
     1561 cggtgaacag ctcctcgccc ttgctcacca ttggcccggg attctcttcg acatcccctg
     1621 cttgtttcaa cagggagaag ttagtggcat cgatggcacc gggcttgcgg gtcatgcacc
     1681 aggtgcgcgg tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt
     1741 agaaggggag gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg
     1801 ccgcctccac tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg
     1861 agacgccgac ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc
     1921 cttccatctg ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga
     1981 tctcggcgaa caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg
     2041 cgccgtcgtc cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt
     2101 cttgcagctc ggtgacccgc tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg
     2161 ggtagtcggc gaacgcggcg gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg
     2221 cgaggcgcac cgtgggcttg tactcggtcc ccatggtggc gttggctgca ggtcgaaagg
     2281 cccggagatg aggaagagga gaacagcgcg gcagacgtgc gcttttgaag cgtgcagaat
     2341 gccgggcctc cggaggacct tcgggcgccc gccccgcccc tgagcccgcc cctgagcccg
     2401 cccccggacc caccccttcc cagcctctga gcccagaaag cgaaggagca aagctgctat
     2461 tggccgctgc cccaaaggcc tacccgcttc cattgctcag cggtgctgtc catctgcacg
     2521 agactagtga gacgtgctac ttccatttgt cacgtcctgc acgacgcgag ctgcggggcg
     2581 ggggggaact tcctgactag gggaggagta gaaggtggcg cgaaggggcc accaaagaac
     2641 ggagccggtt ggcgcctacc ggtggatgtg gaatgtgtgc gaggccagag gccacttgtg
     2701 tagcgccaag tgcccagcgg ggctgctaaa gcgcatgctc cagactgcct tgggaaaagc
     2761 gcctccccta cccggtagaa ttgctagcgt gttcaatact tattccctgt gtcatttcac
     2821 tttatttatt atgactcaac ttgtatactt aaatgttctg atttctttgt atgaattcaa
     2881 tatttggctt gatggctaca tctggtagga attttgtgtc agtagcccac ttagaaatcc
     2941 ccttactgat aaaaatgctg atgtgtcaaa tacttatttt ccccgctgta catatgagag
     3001 acgcaagaca ctgcggatcg agacgcattg cgtggttcga ctggccgtcg ttttacaacg
     3061 aaagggcctc gtgatacgcc tattgactcc agcgtaactg gactgcaatc aactcactgg
     3121 ctcaccttca cgggtgggcc tttcttcggt agaaaatcaa aggatcttct tgagatcctt
     3181 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3241 gtttgccgga tcaagagcta ccaactcttt ttccgaggta actggcttca gcagagcgca
     3301 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3361 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3421 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3481 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3541 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3601 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3661 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcatcgatt
     3721 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cagaaaggcc
     3781 cacccgaagg tgagccaggt gattacattt gggccctcat cagaggtttt caccgtcatc
     3841 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     3901 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     3961 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca cttaccaatg
     4021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4081 actccccgtc gtgtagataa ctacgatgcg ggagggctta ccatctggcc ccagtgctgc
     4141 aatgataccg cgggacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4201 cgggagggcc gagcgcagaa gtgatcctgc aactttatcc gcctccatcc agtctattaa
     4261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4441 cttcggtcct ccgatcgttg ccagaagtaa gttggccgca gtgttatcac tcatggttat
     4501 ggcagcactg cataattctc ttactgtcat gccatccgtg agatgctttt ctgtgactgg
     4561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4681 aaaacgttct tcggggcgta aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4741 gtaacccac
//
