LOCUS       sequence-442192-        5193 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of HAT2_CI Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5193)
  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 5193)
  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-HAT2_CI
FEATURES             Location/Qualifiers
     source          1..5193
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..112
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     121..138
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     122..138
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        638..1137
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(673..696)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1070..1089
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1151..1160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1157..1756
                     /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     1656..1676
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1763..1819
                     /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(1820..1822,1823..1825,1826..2539)
                     /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(1820..1822,1823..1825,1826..2536)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1865..1886)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2126..2145)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2473..2494
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2561..2578)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2567..2791
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3296..3312)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3296..3312)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3324..3341)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3495..4083)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3575..3594)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4254..5045,5046..5114))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLRNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERKRQIAEIGAS
                     LIKHW"
     primer_bind     4877..4896
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5115..5193,1..26))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
       61 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
      121 tgtaaaacga cggccagtcg aaccacgcaa tgcgtctcga tccgcagtgt cttgcgtctc
      181 tactagtata ataattaggg ctgtggattt tcgccttttt ttgttaaccg ttaaccgttc
      241 agttttaacc ggttaaccgg ttaaccgata caagattagc tgttgcttaa aacagcgtag
      301 tgagatacgc tgtgtgtcat aaatggggac ttttaaagcg tggcggcaat ttgtccttga
      361 ttggcatcaa gcgcgtggct gaccggattt atattgcaac acatgtgcgc acacgaggca
      421 acctaaacaa attcttatca tttaaggttt tgacgtgatg catgttacca tggttattta
      481 aagatattgc accatcttta gaatacattc ctctataaat acgtcataat aacttttata
      541 cgatcataaa caccagtcta taacaaaccc acacattttt ttgctgcgtg tcagttatta
      601 aattgtatgc aacaataccg gtgctagcaa ttctaccggg taggggaggc gcttttccca
      661 aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta cacaagtggc
      721 ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc gttctttggt
      781 ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc cgccccgcag
      841 ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct cgtgcagatg
      901 gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca atagcagctt
      961 tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg cgggctcagg
     1021 ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc attctgcacg
     1081 cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg cctttcgacc
     1141 tgcagccaac gccaccatgg ggaccgagta caagcccacg gtgcgcctcg ccacccgcga
     1201 cgacgtcccc cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
     1261 ccacaccgtc gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
     1321 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
     1381 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
     1441 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
     1501 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
     1561 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
     1621 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
     1681 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
     1741 ccgcaagccc ggtgccatcg atgccactaa cttctccctg ttgaaacaag caggggatgt
     1801 cgaagagaat cccgggccaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1861 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1921 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1981 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2041 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2101 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2161 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2221 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2281 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     2341 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2401 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     2461 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2521 ggacgagctg tacaagtaat taattaagct cgctgatcag cctcgactgt gccttctagt
     2581 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2641 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2701 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2761 aggcatgctg gggatgcggt gggctctatg gcttctgagg cgcgccctcg agctatactc
     2821 tgcaaagatt gtaaatagca gttaagattg taaatagcag tttgtaacgt gttgacattt
     2881 taaatggtgt ttggaaatct tcaattgaat gtgaaataca ctctttctat taatttttct
     2941 tttatttttt atgactattc atgacgtggc aggttcgtgg ttgcgcaaga gaaaaaatag
     3001 ttcacctgag gtattaacaa tcgagattga taacgcaatc gctgttagag gagtgctcga
     3061 tcaaagccac agcgtcgctt atcgcgaagc tacaatcaac gaacgcggac gcgcgtttcg
     3121 tgtaaataat acattgctta atataagtgt gtgcggatga cgtcataatt cggttaacga
     3181 ctcgaacttt ttgttagcgg ttaaccgaat agtttcggtt aaccggttaa ccgttaactt
     3241 ttccacagcc ctaataataa tgggcccaga gacggagtca ctgccaaccg agacggtcat
     3301 agctgtttcc tgtgtgccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3361 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tacccacaga atcaggggat
     3421 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3481 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3541 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3601 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3661 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3721 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3781 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3841 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3901 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3961 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4021 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4081 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4141 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4201 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4261 tgcttaatca gtgaggcacc tatctcagcg atctgtctct ttcgttcatc catagttgcc
     4321 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4381 gcaataatac cgcgggaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4441 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     4501 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     4561 gccatcgcta caggcatcgt ggtatcacgc tcgtcgtttg gtatggcttc attcagctcc
     4621 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgcgcaaaaa agcggttagc
     4681 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg ccgtgttatc actcatggtt
     4741 atggcagcac tacataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     4801 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     4861 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     4921 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     4981 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5041 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5101 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5161 ctcatgagcg gatacatatt tgaatgtatt tag
//
