LOCUS       sequence-443564-        5131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-19B_Croc Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5131)
  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 5131)
  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-hAT-19B_Croc
FEATURES             Location/Qualifiers
     source          1..5131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      12..39
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(51..638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(131..150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(660..689)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     751..768
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     752..768
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1196..1695
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1231..1254)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1628..1647
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1709..1718
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1715..2314
                     /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     2214..2234
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2321..2377
                     /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(2378..2380,2381..2383,2384..3097)
                     /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(2378..2380,2381..2383,2384..3094)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2423..2444)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2684..2703)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3031..3052
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3119..3136)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3125..3349
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3835..3851)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3835..3851)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3863..3880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      3932..3963
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        3964..4066
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(4067..4135,4136..4927)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGHRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAELGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4285..4304)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             4928..5119
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tgtaatcacc tggctcacct tcgggtgggc ctttctgcgt tgctggcgtt tttccatagg
       61 ctccgccccc ctgacgagca tcacaaaaat cgatgctcaa gtcagaggtg gcgaaacccg
      121 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      181 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      241 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      301 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
      361 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
      421 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
      481 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac ctcggaaaaa
      541 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
      661 gaagaaaggc ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
      721 gtcaaaaggc gtatcacgag gccctttcgt tgtaaaacga cggccagtcg aaccacgcaa
      781 tgcgtctcga tccgcagtgt cttgcgtctc tcaattgctc ataaataggg acctaccgaa
      841 atcgcgattt cgcgtttttc gcggaaaacg cgaaatcggg cgttctccgc gaaaaacggc
      901 gggctccgcg attttggcgg gctcgctgca gaaaaaaaaa cccaacttac cggcaagcag
      961 agaaaagggc gggaagccct gcagtcctga agcacggggg gtggggagag gaggggggag
     1021 agggagcaaa tcagaggctg agcctgtgac gtaggctcag tacaggagcc agtccggggc
     1081 cagtcaggga agcgagaggg ttttggcgcc atcttttaag cagccagagg catgctgctg
     1141 cagcatgctg cagcctgagg aggattgagc agggaccggt gctagcaatt ctaccgggta
     1201 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     1261 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     1321 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     1381 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     1441 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     1501 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     1561 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     1621 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     1681 tctccgggcc tttcgacctg cagccaacgc caccatgggg accgagtaca agcccacggt
     1741 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     1801 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     1861 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     1921 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     1981 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     2041 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     2101 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     2161 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     2221 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     2281 gcgcacctgg tgcatgaccc gcaagcccgg tgccatcgat gccactaact tctccctgtt
     2341 gaaacaagca ggggatgtcg aagagaatcc cgggccaatg gtgagcaagg gcgaggagct
     2401 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     2461 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     2521 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2581 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2641 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     2701 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2761 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     2821 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     2881 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     2941 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     3001 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     3061 cgggatcact ctcggcatgg acgagctgta caagtaatta attaagctcg ctgatcagcc
     3121 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3181 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3241 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3301 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3361 cgccctcgag catgaattgc ctgtctcaag gttagcattg agtaaactcc tttttacaac
     3421 gtcctttaat aaagtatatg tagatggtat tagttacaat tcatgttgtg tatagcttta
     3481 atttcataag acatttcagt ttagagttta attaaatgct tagattgctt aataacgcgt
     3541 caattaatgg cttccgcggg cttaatcccc tgattaagcc cgcggaagcc attaagggaa
     3601 acattattaa gcaattaagc cgcggtttcg ggccatctgg caatctgcgg ggaggggcgg
     3661 gactgccggg gcccctcggc caatcagagg cgtggggggg ccccgccgcg atcagcccgc
     3721 gaaaatggcg gccggccccg ggatctgccc gcgaaatcgg ccggccgcct cctcgagttc
     3781 ggtagacccc tactcataaa gggcccagag acggagtcac tgccaaccga gacggtcata
     3841 gctgtttcct gtgtgccact tcctcgctca ctgactcgct gcgctcggtc gttactgacc
     3901 atttaaatca tacctgacct ccatagcaga aagtcaaaag cctccgaccg gaggcttttg
     3961 acttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
     4021 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga gtattcaaca
     4081 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4141 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4201 cgaactggat ctcaacagcg gtaagatcct tgagagttta cgccccgaag aacgttttcc
     4261 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4321 gcaagagcaa ctcggtcacc gcatacacta ttctcagaat gacttggttg agtactcacc
     4381 agtcacagaa aagcatctca cggatggcat gacagtaaga gaattatgca gtgctgccat
     4441 aaccatgagt gataacactg cggccaactt acttctggca acgatcggag gaccgaagga
     4501 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4561 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4621 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4681 aatagactgg atggaggcgg ataaagttgc aggatcactt ctgcgctcgg ccctcccggc
     4741 tggctggttt attgctgata aatctggagc cggtgagcgt gggtcccgcg gtatcattgc
     4801 agaactgggg ccagatggta agccctcccg catcgtagtt atctacacga cggggagtca
     4861 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4921 ttggtaagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     4981 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     5041 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     5101 tgacggtgaa aacctctgat gagggcccaa a
//
