LOCUS       sequence-443302-        5202 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-4_Crp Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5202)
  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 5202)
  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-4_Crp
FEATURES             Location/Qualifiers
     source          1..5202
                     /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        819..1318
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(854..877)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     1251..1270
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      1332..1341
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1338..1937
                     /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     1837..1857
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1944..2000
                     /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(2001..2003,2004..2006,2007..2720)
                     /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(2001..2003,2004..2006,2007..2717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2046..2067)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2307..2326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2654..2675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2742..2759)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2748..2972
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3305..3321)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3305..3321)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3333..3350)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3504..4092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3584..3603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4263..5054,5055..5123))
                     /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     4886..4905
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5124..5202,1..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 tcaattgcta tatagcaggg cccaagcatg gggggggggg aagggtacac tttaccaggg
      241 cccggagatc caggggggcc cagcccaggc tgcaacccta ttaccatgag gaggaaaaaa
      301 cgaaggctaa attgtaggtc atgcttgagg atatgtatgg aaagttatgt actacctata
      361 tggggtctcc actgtgagat ttgctggagc caatcagaag cagaacaagc aacattgtga
      421 acagtgagtt gctggccagc ccagggccct tctgtagaac agagagagtg tgcttgtgca
      481 gttaattcta ggacttgtag tttgagagac agagggattt atgcttccct atgtgacatg
      541 ctctctcaag aactacaagt cccagagtcc cttgtgcata tagatggaga ggggagggac
      601 aaagcaccat ataaccaggg acattgatcc cagggacagt ggaagccaca gtgttgctgg
      661 taggatttac aggcccaaaa gtgagtaatg agttaggcct ggtggtctaa acattgcaca
      721 gagaagaact gaaacaggga tagggttagt gcctggaagg aaaggacatt tctttctctc
      781 tctctcacac acacaccacc ggtgctagca attctaccgg gtaggggagg cgcttttccc
      841 aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg
      901 cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg
      961 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca
     1021 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat
     1081 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct
     1141 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag
     1201 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac
     1261 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac
     1321 ctgcagccaa cgccaccatg gggaccgagt acaagcccac ggtgcgcctc gccacccgcg
     1381 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     1441 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     1501 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     1561 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     1621 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     1681 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     1741 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     1801 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     1861 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     1921 cccgcaagcc cggtgccatc gatgccacta acttctccct gttgaaacaa gcaggggatg
     1981 tcgaagagaa tcccgggcca atggtgagca agggcgagga gctgttcacc ggggtggtgc
     2041 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     2101 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     2161 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     2221 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     2281 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     2341 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     2401 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     2461 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     2521 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     2581 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     2641 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     2701 tggacgagct gtacaagtaa ttaattaagc tcgctgatca gcctcgactg tgccttctag
     2761 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2821 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2881 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2941 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcgcgccctc gagagtgaat
     3001 tggtctattt ataaaatgtt tgcagccatg ttggtccatt agaacaaaaa aaatcaaaac
     3061 atcgactgga taataccatt attaggacca acaaccaaaa agttacaaaa tagtcagcaa
     3121 tgtagaatgt aatttcagaa ttaaaataga aaattttctt tcatgattca tgagatttta
     3181 tgtatgtggg atggtgtgtg tgtgggaggg ggcccgcctt acaagtttgt actgggccca
     3241 aagaaatgat gcttgggccc tgctatatag gggcccagag acggagtcac tgccaaccga
     3301 gacggtcata gctgtttcct gtgtgccgct tcctcgctca ctgactcgct gcgctcggtc
     3361 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt acccacagaa
     3421 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3661 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3721 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3781 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3841 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3901 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     3961 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4021 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4081 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4141 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4201 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4261 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctctt tcgttcatcc
     4321 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4381 cccagtgctg caataatacc gcgggaccca cgctcaccgg ctccagattt atcagcaata
     4441 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4501 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4561 aacgttgttg ccatcgctac aggcatcgtg gtatcacgct cgtcgtttgg tatggcttca
     4621 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gcgcaaaaaa
     4681 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc cgtgttatca
     4741 ctcatggtta tggcagcact acataattct cttactgtca tgccatccgt aagatgcttt
     4801 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4861 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4921 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     4981 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5041 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5101 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5161 ggttattgtc tcatgagcgg atacatattt gaatgtattt ag
//
