LOCUS       sequence-461270-        4111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bigMamAct is evolution of biGBac for mammalian cells. psBIG plasmids
            are recipient of multi-assembly step by Gibson cloning .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4111)
  AUTHORS   Pelosse M, Marcia M
  TITLE     biGMamAct: efficient CRISPR/Cas9-mediated docking of large 
            functional DNA cargoes at the ACTB locus
  JOURNAL   Synthetic Biology 2025.02.13
REFERENCE   2  (bases 1 to 4111)
  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"; journalName: "Synthetic 
            Biology 2025.02.13"
COMMENT     Sequence Label: psBIG2abcd
FEATURES             Location/Qualifiers
     source          1..4111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        303..405
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             406..1065
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(472..491)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1423..2011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1912..1931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2165..2182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2316..2540
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2607..3140)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3329..3334,3335..3351,3352..3357))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(3753..3786)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  3864..4029
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
       61 ctgaatatca acgcttattt gcagcctgaa ctgaacagga gggacagctg atagaaacag
      121 aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt ggcagcatca
      181 cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc agaataaata
      241 aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg aaaatgagac
      301 gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc
      361 gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
      421 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
      481 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
      541 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
      601 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
      661 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
      721 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
      781 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
      841 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
      901 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
      961 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     1021 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt ttaaggcagt
     1081 tattggtgcc cttaaacgcc tggttgctac gcctgaataa gtgataataa gcggatgaat
     1141 ggcagaaatt cgaaagcaaa ttcgacccgg tcgtcggttc agggcagggt cgttaaatag
     1201 ccgcttatgt ctattgctgg tttaccggtt tattgactac cggaagcagt gtgaccgtgt
     1261 gcttctcaaa tgcctgaggc cagtttgctc aggctctccc cgtggaggta ataattgacg
     1321 atatgatcat ttattctgcc tcccagagcc tgatggcgaa tgggacgcgc ccttttcgtt
     1381 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1441 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1501 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1561 aaatactatt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1621 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1681 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1741 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1801 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1861 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1921 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1981 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     2041 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     2101 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2161 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     2221 gcatctgtgc ggtatttcac accgcataga ccagccgcgt aacctggcaa aatcggttac
     2281 ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg gcggacaata aagtcttaaa
     2341 ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa actagacaga atagttgtaa
     2401 actgaaatca gtccagttat gctgtgaaaa agcatactgg acttttgtta tggctaaagc
     2461 aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc gtggccaagg
     2521 gcatggtaaa gactatattc gcggcgttgt gacaatttac cgaacaactc cgcggccggg
     2581 aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat atcaaagtgc
     2641 atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc gtcaccgtaa
     2701 tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt gaggagattg
     2761 atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag atcatagata
     2821 tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag agcgccaaca
     2881 accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag caagttcccg
     2941 aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa ctcacgaccg
     3001 aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct acatgtgcga
     3061 atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg ctgcgtaaca
     3121 tcgttgctgc tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta
     3181 acgcgcttgc tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc
     3241 catgaaaacc gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt
     3301 gcgtgagcgc atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg
     3361 ggtttgtgcc ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga
     3421 agtcgaggca tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg
     3481 tcaggcattg gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctt
     3541 gcttcaggag atcggtagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga
     3601 tgaagtggtt cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc
     3661 tagctatagt tctagtggtt ggctacagct ttgtttgtac tatcaacagg ttgaactgct
     3721 gatcaacaga tcctctacgc ggccgcggta ccataacttc gtatagcata cattatacga
     3781 agttatctgg cctaggttaa ttaaacgctc tatggtctaa agtttaaacg gttcacatag
     3841 cttagtttca atcaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt
     3901 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     3961 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     4021 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc
     4081 catgtgacaa accgtcatct tcggctactt t
//
