LOCUS       sequence-461276-        4107 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 4107)
  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 4107)
  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: psBIG2ab
FEATURES             Location/Qualifiers
     source          1..4107
                     /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      1419..2007
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1908..1927
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2161..2178
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2312..2536
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2603..3136)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3325..3330,3331..3347,3348..3353))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(3749..3782)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  3860..4025
                     /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 cccgttccac
     1381 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     1441 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     1501 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     1561 actattcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     1621 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     1681 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     1741 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     1801 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     1861 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     1921 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     1981 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2041 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2101 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     2161 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     2221 ctgtgcggta tttcacaccg catagaccag ccgcgtaacc tggcaaaatc ggttacggtt
     2281 gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga
     2341 acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag ttgtaaactg
     2401 aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc taaagcaaac
     2461 tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg ccaagggcat
     2521 ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg gccgggaagc
     2581 cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca aagtgcatca
     2641 cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca ccgtaatctg
     2701 cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg agattgatga
     2761 gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca tagatataga
     2821 tctcactacg cggctgctca aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg
     2881 cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag ttcccgaggt
     2941 aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca cgaccgaaaa
     3001 gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat gtgcgaatga
     3061 tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc gtaacatcgt
     3121 tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac ggcgtaacgc
     3181 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     3241 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     3301 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     3361 tgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     3421 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     3481 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gcccttgctt
     3541 caggagatcg gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac cccggatgaa
     3601 gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca ggactctagc
     3661 tatagttcta gtggttggct acagctttgt ttgtactatc aacaggttga actgctgatc
     3721 aacagatcct ctacgcggcc gcggtaccat aacttcgtat agcatacatt atacgaagtt
     3781 atctggccta ggttaattaa acgctctatg gtctaaagtt taaactggat actattgcac
     3841 gtttcaatca ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt
     3901 tttaattttc gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc
     3961 cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc
     4021 ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg
     4081 tgacaaaccg tcatcttcgg ctacttt
//
