LOCUS       sequence-443457-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of hAT-2_AG Transposon.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  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 7374)
  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-2_AG
FEATURES             Location/Qualifiers
     source          1..7374
                     /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     724..742
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     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        3534..4033
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(3569..3592)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3966..3985
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      4047..4056
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4053..4652
                     /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     4552..4572
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             4659..4715
                     /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(4716..4718,4719..4721,4722..5435)
                     /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(4716..4718,4719..4721,4722..5432)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4761..4782)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5022..5041)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5369..5390
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5457..5474)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5463..5687
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6079..6095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6079..6095)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6107..6124)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      6176..6207
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        6208..6310
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(6311..6379,6380..7171)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAELGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6529..6548)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
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 ggaaaacaaa ccaccgctgg tagcggtggt ttttttgttt
      601 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg attttctacc
      661 gaagaaaggt ccacccgtga aggtgagcca gtgagttgat tgcagtccag ttacgctgga
      721 gtcaataggc gtatcacgag gccctttcgt tgtaaaacga cggccagtcg aaccacgcaa
      781 tgcgtctcga tccgcagtgt cttgcgtctc tctgcaggta tggactaggc atgggctaat
      841 tgtgcgagac ctatgccgca cactcagttc actcaagtgt gcggctgacc accgcacgcg
      901 aaaaagtgta cgggtaggtc ttccgcacgc ataaggaatg acttcggttg tgactggttg
      961 aacgaaaaaa aaaacattaa ttttagtcga ctgtgttcaa cagtcaaata aaaaatcgat
     1021 cacactctga gcgagaaaga atacgcaaat tgttagcgta tttagttggg tcagtcgcac
     1081 taaaagaact cctgtccaag actgacatgg aacgagaaat acactcatac ccgttgggtc
     1141 agtcgcacac tgccgcacgc gtgttcagtt tggtcagtcg cacacacagc tgcacaaaaa
     1201 gaaattctgt cgcacactgc cgcacgtttg ctcagttcgt tcattcgcac acagccgcac
     1261 aaaaagaact tctggcgcac actgccgcac gcgtgttcag ttgggtcagt cgcacacagc
     1321 cgcacaaaaa gaacttctgc cgcacactgc cgcacgtttg ctcaattcgt tcattcgcac
     1381 acagccgcac aaaaagaact tctgccgcac gctgccgcac gtgaggtcag ttgggtcagt
     1441 cgcacacagc cgcacaaaaa gaacttctgc cgcacactgc cgcacgtttg ctcaattcgt
     1501 tcattcgcac acagccgcac aaaaagaact tctgccgcac gctgccgcac gtgaggtcag
     1561 ttgggtcagt cgcacacagc cgcacaaaaa taactcacac tgactacgct agtaatatgc
     1621 tcgttctctc tgcgctgtcg gatcagtcgc acacagtcat acaaagaact gctaccgcac
     1681 actgtcctga tgaaacacag ggtttgatgt ctttttggtt gtttcttcat tttttggatt
     1741 tatctcacat tttattgcat gcttcccaaa ttattttgga ctcattccac aatttattat
     1801 atgtttctca tataattttg tattcgtttt gtgatttatt cttaccatcc cagttgtttt
     1861 gaaacaaata attgaacaaa ttttatgaaa cgttaaataa tcgtgatcca taaaccctgt
     1921 aataggagac aaaattggta tcaatagcat cttcttcctt ctcattttct gctgattgtt
     1981 ttttacgtag caccagacat tcttagcgtg gggaactgtg tggtattgat atgtatcgaa
     2041 tgtaagagca aggagggaaa aacgtacact aggtattcgc tagttgaatg atttgaaatg
     2101 tggatcattt ttttaagttg catggtcgat gtgagattta taaatcgatg atttttaatc
     2161 ttgaaatatt caccacatgc aaaatcaaaa aatctccaaa tctttatatt cttttgcttt
     2221 ttatcattta aaaaaatatg ccaattcaaa atgcaataca tgtcacacat tacaatttca
     2281 attagcgagc tcaaactgta ccatcccacg atgttttatt cgaacaccat tactagtgag
     2341 gcatgctttt cgcacactaa gttgatgagt gcaatatgta tataaaagaa cgtcaagtat
     2401 tcggaaagtg gcttacccat tatgaatgag tccctcgttt gttggtgttg tatgaataat
     2461 attgcagtag gtaacaaagt gaaatagtat tgaagtgcgt ttccggtatc agagttttag
     2521 ctttgcattg tcgttgcgat tcctattttg gttgtgtatc gaatcagtcg cattaaagga
     2581 aagtacccag tttgaaaatt cataacgcgt aaatactttt tctgtggaag aatcggcagg
     2641 gataacctga taactaatta aaggtaagtt gctttattat ttttaacttt taacaagatg
     2701 agaaaaaatg acaaatcgtc atttattctt ttttttattt cataaaaaaa atcctagttc
     2761 agtggtcgag aaagtacggc aatccgttta cgatttttgc cagcattttc actccttaac
     2821 agccccttgc tattgtagga gtattcaaaa tctcccacag tttcacgcct tcatctgaaa
     2881 atccggtttg gaaatttact gacatttgta tagaatttag ctgtctatgg cgtcaagtta
     2941 accgacgttt tttttctagt tcagtggtca gcaaactttg tggtgaaaag agccaaattc
     3001 tatgaaaatg ttggtagatt tcgatatgaa gagccaaatt tgaaaaccaa gcatacaatt
     3061 gagttaacca ctgtcctcat ttggaccttc cttccacaaa tgacattcaa agccaaaact
     3121 attcaatgtt ttggtcacac cagtgacaac tatgatcgac tgacttttca tgatgactga
     3181 gtaactttta atcgaagtaa tagatttaat cattatagta aaatttacat catcaattaa
     3241 aaataatgat ctacaattta tcaataatta aagtttcctc aaaataccaa agcacagtga
     3301 tagtattacg tttattttat ttgaataata ttgctataat tataatttac aacaattgct
     3361 gacgggactg gaactatacc aaaacgttct ttaattaaaa aaaaactgtt cgttttagca
     3421 gtcatactgt taaagccatt taggctgtta gataattgaa caaatattta aaattttatt
     3481 tttgattaat gttgtaatat gttttagtta agaccggtgc tagcaattct accgggtagg
     3541 ggaggcgctt ttcccaaggc agtctggagc atgcgcttta gcagccccgc tgggcacttg
     3601 gcgctacaca agtggcctct ggcctcgcac acattccaca tccaccggta ggcgccaacc
     3661 ggctccgttc tttggtggcc ccttcgcgcc accttctact cctcccctag tcaggaagtt
     3721 cccccccgcc ccgcagctcg cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac
     3781 tagtctcgtg cagatggaca gcaccgctga gcaatggaag cgggtaggcc tttggggcag
     3841 cggccaatag cagctttgct ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc
     3901 cgggggcggg ctcaggggcg ggctcagggg cggggcgggc gcccgaaggt cctccggagg
     3961 cccggcattc tgcacgcttc aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc
     4021 tccgggcctt tcgacctgca gccaacgcca ccatggggac cgagtacaag cccacggtgc
     4081 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     4141 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     4201 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     4261 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     4321 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     4381 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     4441 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     4501 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     4561 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     4621 gcacctggtg catgacccgc aagcccggtg ccatcgatgc cactaacttc tccctgttga
     4681 aacaagcagg ggatgtcgaa gagaatcccg ggccaatggt gagcaagggc gaggagctgt
     4741 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     4801 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     4861 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     4921 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     4981 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     5041 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     5101 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     5161 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     5221 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     5281 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     5341 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     5401 ggatcactct cggcatggac gagctgtaca agtaattaat taagctcgct gatcagcctc
     5461 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     5521 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     5581 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     5641 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgcg
     5701 ccctcgagaa aattatttag tttttggatt gaatttaggt aagcctaaga atgaaataaa
     5761 ttgttagtgt gtatgatata ttttttataa cttttctctt aaactaaaaa tttataaatt
     5821 agaattgtaa caccatggac taaacatctc aacaaataca taaaagtaaa gttttgttcg
     5881 agcatccgca ttatttattt aaaataaaag acagttcaaa atttaattct ttgtgtgacc
     5941 gacagtgcga ctgacctacc gcacgcgttc agttcctcat actgaacgaa ctacaccgca
     6001 cgcgttcact gaaaagaatc aaattgccca agcctagtat ggacgggccc agagacggag
     6061 tcactgccaa ccgagacggt catagctgtt tcctgtgtgc cgcttcctcg ctcactgact
     6121 cgctgcgctc ggtcgttact gaccatttaa atcatacctg acctccatag cagaaagtca
     6181 aaagcctccg accggaggct tttgacttga tcggcacgta agaggttcca actttcacca
     6241 taatgaaata agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa
     6301 ggaagctaaa atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     6361 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     6421 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     6481 tttacgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     6541 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     6601 gaatgacttg gttgagtact caccagtcac agaaaagcat ctcacggatg gcatgacagt
     6661 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     6721 ggcaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     6781 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6841 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6901 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggatc
     6961 acttctgcgc tcggccctcc cggctggctg gtttattgct gataaatctg gagccggtga
     7021 gcgtgggtcc cgcggtatca ttgcagaact ggggccagat ggtaagccct cccgcatcgt
     7081 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     7141 gataggtgcc tcactgatta agcattggta agtgaccaaa caggaaaaaa ccgcccttaa
     7201 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     7261 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     7321 cagctgcctc gcgcgtttcg gtgatgacgg tgaaacctct gatgagggcc caaa
//
