LOCUS       sequence-443592-        5460 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 5460)
  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 5460)
  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-Harbinger-3-AMi
FEATURES             Location/Qualifiers
     source          1..5460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     212..232
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             319..375
                     /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(376..378,379..381,382..1095)
                     /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(376..378,379..381,382..1092)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(421..442)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(682..701)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1029..1050
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1117..1134)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1123..1347
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2167..2183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2167..2183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2195..2212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2264..2295
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        2296..2398
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             join(2399..2467,2468..3259)
                     /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(2617..2636)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             3260..3451
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      3475..3502
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(3514..4101)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3594..3613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4123..4152)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     primer_bind     4187..4205
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     4214..4231
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4215..4231
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4654..5153
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4689..4712)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     5086..5105
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      5167..5176
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5173..5460,1..312)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
ORIGIN
        1 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
       61 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
      121 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
      181 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
      241 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
      301 aagcccggtg ccatcgatgc cactaacttc tccctgttga aacaagcagg ggatgtcgaa
      361 gagaatcccg ggccaatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      421 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      481 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      541 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
      601 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
      661 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
      721 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
      781 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
      841 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
      901 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
      961 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1021 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1081 gagctgtaca agtaattaat taagctcgct gatcagcctc gactgtgcct tctagttgcc
     1141 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1201 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1261 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1321 atgctgggga tgcggtgggc tctatggctt ctgaggcgcg ccctcgagcg gggctcccgc
     1381 gccggctctc cgggcctccg cgcggctgtc cctgtgctgc cgggcctggg cagctgcctg
     1441 cagcgcccag cgcaggggca cggcacggga ctggcagccg cgcggcagag gcaatgcccc
     1501 gcagcacctg caggctccgg gcaggctgct gggggtgggc tgcgggcagc tggagctggc
     1561 cccggcggcc aggagctgcc tgcagctggg cctgggggct ggcaggcctg caggagcctg
     1621 cagcagcctg caggcctgga cggcgcgggc ctggcagccg cgcggcactg cggtgccagg
     1681 cagcggcgcc ggggtgcagg caggctggcg cggccggcct gcggcctgcc cagggggaca
     1741 caggggcctg gagtggcctc aggctccctg ctggtgtgcc ccagggctcg aaatgcctgc
     1801 aggcttttaa aggcctgagg caggggctgc catagaggca gcccagccct ggggcaggag
     1861 aggctcccag cccggagccc tggggctagc ctcaggccgg ccccctagtg gcagggctga
     1921 cctgggagca aatttgctcc cgggtcgcgt ctacacgtgc gttactgcgc agtaactaat
     1981 cgcatttgca ggtagcaaat ttactcgcga ttagcgacgt ttactgcgca gtaagcgtgc
     2041 gcacgtgtag acgcgcgcgc ttactgcgca gtaaactacg ctactgcgca gttaagcgtc
     2101 tcgtgtagac gcgcccactg tgattcttct gtggtaccag agacggagtc actgccaacc
     2161 gagacggtca tagctgtttc ctgtgtgccg cttcctcgct cactgactcg ctgcgctcgg
     2221 tcgttactga ccatttaaat catacctgac ctccatagca gaaagtcaaa agcctccgac
     2281 cggaggcttt tgacttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag
     2341 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat
     2401 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2461 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2521 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt tacgccccga
     2581 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2641 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2701 tgagtactca ccagtcacag aaaagcatct cacggatggc atgacagtaa gagaattatg
     2761 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctgg caacgatcgg
     2821 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2881 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2941 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3001 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggatcac ttctgcgctc
     3061 ggccctcccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtcccg
     3121 cggtatcatt gcagaactgg ggccagatgg taagccctcc cgcatcgtag ttatctacac
     3181 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     3241 actgattaag cattggtaag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt
     3301 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca
     3361 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc
     3421 gcgtttcggt gatgacggtg aaaacctctg atgagggccc aaatgtaatc acctggctca
     3481 ccttcgggtg ggcctttctg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3541 gcatcacaaa aatcgatgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3601 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3661 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3721 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3781 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3841 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3901 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3961 atttggtatc tgcgctctgc tgaagccagt tacctcggaa aaagagttgg tagctcttga
     4021 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     4081 cgcagaaaaa aaggatctca agaagatcct ttgattttct accgaagaaa ggcccacccg
     4141 tgaaggtgag ccagtgagtt gattgcagtc cagttacgct ggagtcaata ggcgtatcac
     4201 gaggcccttt cgttgtaaaa cgacggccag tcgaaccacg caatgcgtct cgatccgcag
     4261 tgtcttgcgt ctctaagctt atatataaga cacagtgggc gcgtctacac gtgcatttga
     4321 tgcggcacca gtttactcgc gagtaaacta ctcgcgagta aacggtcccg ggcaggcgtc
     4381 tacacgtgca gggaagcggg agcagatttg ctgccaatct gctcccgctt cccggggccc
     4441 tgcaatgggc ccctgccgcc accgggggga gctccaggct ccggctgcag ctgccagcag
     4501 ccggcagccc tcttcaaaag ccagccccgc tctctgctcc ctggccagcc acttccctgg
     4561 ccaggagcag cgtgcacagc gcaggggcag cacgcggcag ctctctcctc gccgcctctg
     4621 cctccccgcg ccaccggtgc tagcaattct accgggtagg ggaggcgctt ttcccaaggc
     4681 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
     4741 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
     4801 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
     4861 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
     4921 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
     4981 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
     5041 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
     5101 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
     5161 gccaacgcca ccatggggac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     5221 gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     5281 accgtcgacc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     5341 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5401 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
//
