LOCUS       sequence-443570-        6347 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-5_Croc Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6347)
  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 6347)
  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-5_Croc
FEATURES             Location/Qualifiers
     source          1..6347
                     /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        1619..2118
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1654..1677)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2051..2070
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      2132..2141
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2138..2737
                     /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     2637..2657
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2744..2800
                     /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(2801..2803,2804..2806,2807..3520)
                     /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(2801..2803,2804..2806,2807..3517)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2846..2867)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3107..3126)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3454..3475
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3542..3559)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3548..3772
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4450..4466)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4450..4466)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4478..4495)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4649..5237)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4729..4748)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5408..6199,6200..6268))
                     /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     6031..6050
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6269..6347,1..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 tcaattgctt taaagcaggg tggataaaaa tcaacgattt ttaaaaaaaa aattaaaaat
      241 cggatttttt tatttaatcg tattattttt ttttgaagat gctttaaaaa aacctatata
      301 aagatagttt taatttaaga tacgttaaag ctcaaagata tcatcatgga atagggatta
      361 taacttctaa ttatatacta tttgagacaa tatattcgtg taaccttttt agaaaagttt
      421 tataaatagg tcacaacagg ccatggacta tattaggggc ccaatcttat ggggttccca
      481 gaggctcctc tatagattag taaagattaa agaaaaccac tctacccaat gggactcagt
      541 gctcagtcta gaagatccca ttaagcatct ctgtgatgct tagtttcagt tctcaaactg
      601 tggattttgt gtctccagag ataacatcct tgttaacagc agtatatgga gatgagagat
      661 aacagacctg attacctacc catcagccct attgtctctc tgcaagtttg tgtacacaga
      721 gtcaagccct tacctctctc taaaagtgca aagtttcaag aagttaaatg aatagaggat
      781 tgttgggggt ggagtagatc tgggcaagga ggagtctgga gataaatgca aggagggagg
      841 ggcagtagaa acaaaagaga aacattttga gcagaatatt ccagaagtct tgagggataa
      901 gtttctgagt atagcctctc cattgatttg agatctagtg agggaatggt atggtagaag
      961 ggaaaaccta tcatggcagc aggcttgcca ggccataaaa gagaccctgt ttgggaatat
     1021 tttaatgaag ttcctctacc tgtgggtaag acaggcatgt gtgcaaaatg caaacagtgc
     1081 aacaaagaaa tgcaaggcct ggttgcccga atgaaacaac atcatgagaa gtgctccttc
     1141 tcaggaggaa gctgctttga agatgatgaa tggaacatgt ctgaacatgc aggatctcca
     1201 ggttagtaaa ctttcttatt tcatacttct ttcttaagga ctgcctgtct tccctgtcca
     1261 tatcttttgg gtggacttct ggactacttg aattctcata aaaatgtttg agcaaaaata
     1321 tagttgttac tttatggaac tatcatattt ttaatgcagt tgtgatataa aataatggct
     1381 gaaataggca gatcttcctt ttacagtttc acttctaaag tagtactgag tgcaatgagt
     1441 aatactaaat gagcagtact gtagtgactg cattgacttc ttttgttcag gaggatccat
     1501 cctcaacata caggattctg actatccatc ttcaacatca tcactgtcta cagtttcaga
     1561 gttatctgct aatgatacca gtgttccagt caaatgtacc ggtgctagca attctaccgg
     1621 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc
     1681 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc
     1741 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg
     1801 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt
     1861 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg
     1921 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt
     1981 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc
     2041 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc
     2101 tcatctccgg gcctttcgac ctgcagccaa cgccaccatg gggaccgagt acaagcccac
     2161 ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt
     2221 cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg agcgggtcac
     2281 cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc
     2341 ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt
     2401 gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca
     2461 acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac
     2521 cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg
     2581 agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa
     2641 cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg
     2701 accgcgcacc tggtgcatga cccgcaagcc cggtgccatc gatgccacta acttctccct
     2761 gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca atggtgagca agggcgagga
     2821 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2881 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2941 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3001 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     3061 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     3121 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     3181 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     3241 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     3301 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     3361 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     3421 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     3481 cgccgggatc actctcggca tggacgagct gtacaagtaa ttaattaagc tcgctgatca
     3541 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     3601 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     3661 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3721 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3781 gcgcgccctc gaggtttgct gcataaccca gtattttaag tttctcatgt tgacctggct
     3841 gaaataatct atttaatttt tgttaaaaca catttcacat ttataactaa aatagttaaa
     3901 aattcaaaaa tacatatgct tgttttgtta aaataattat acatgtgttg ttgaagaaaa
     3961 atatccaaaa tacataagct tgttgtttta gttaaataaa acaatttaaa tgtctgttgt
     4021 ttttggtgac ttgttgttgt cctcctaata ctggcaaaac ccccatagtg aattggagat
     4081 agttcacctc ccaatgactt cacaagtatc tgcttcaatt agctttggta aataaaataa
     4141 ccaaatcata attcagtttt ctgatgtagc tgcaaaatta atctgaaaaa aatatcaaaa
     4201 taaatcactg ctttaaaatg tatagtgtgt accttctaaa aatgaaacct acatctatct
     4261 ctgagttgtg aataatatgt attaaggtta tatcaaacaa caagaatgca ctttttgtag
     4321 aaaacaatga ttaaatcgag gcttcctgac cagtgattta aatcgtgatt taaatcaaat
     4381 catttgattt aaatcaaatc caccctgctt taaaggggcc cagagacgga gtcactgcca
     4441 accgagacgg tcatagctgt ttcctgtgtg ccgcttcctc gctcactgac tcgctgcgct
     4501 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttaccca
     4561 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4621 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4681 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4741 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4801 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4861 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4921 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4981 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5041 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5101 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5161 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5221 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5281 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5341 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5401 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctctttcgtt
     5461 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5521 ctggccccag tgctgcaata ataccgcggg acccacgctc accggctcca gatttatcag
     5581 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5641 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5701 tgcgcaacgt tgttgccatc gctacaggca tcgtggtatc acgctcgtcg tttggtatgg
     5761 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgcgca
     5821 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgccgtgt
     5881 tatcactcat ggttatggca gcactacata attctcttac tgtcatgcca tccgtaagat
     5941 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6001 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6061 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6121 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6181 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6241 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6301 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttag
//
