LOCUS       sequence-461278-        4114 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 4114)
  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 4114)
  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: psBIG2abc
FEATURES             Location/Qualifiers
     source          1..4114
                     /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      1426..2014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1915..1934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2168..2185
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2319..2543
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2610..3143)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3332..3337,3338..3354,3355..3360))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(3756..3789)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     mobile_element  3867..4032
                     /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 ccgagttttc
     1381 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1441 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1501 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1561 accaaatact attcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1621 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1681 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1741 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1801 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1861 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1921 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1981 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2041 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2101 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2161 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct
     2221 tacgcatctg tgcggtattt cacaccgcat agaccagccg cgtaacctgg caaaatcggt
     2281 tacggttgag taataaatgg atgccctgcg taagcgggtg tgggcggaca ataaagtctt
     2341 aaactgaaca aaatagatct aaactatgac aataaagtct taaactagac agaatagttg
     2401 taaactgaaa tcagtccagt tatgctgtga aaaagcatac tggacttttg ttatggctaa
     2461 agcaaactct tcattttctg aagtgcaaat tgcccgtcgt attaaagagg ggcgtggcca
     2521 agggcatggt aaagactata ttcgcggcgt tgtgacaatt taccgaacaa ctccgcggcc
     2581 gggaagccga tctcggcttg aacgaattgt taggtggcgg tacttgggtc gatatcaaag
     2641 tgcatcactt cttcccgtat gcccaacttt gtatagagag ccactgcggg atcgtcaccg
     2701 taatctgctt gcacgtagat cacataagca ccaagcgcgt tggcctcatg cttgaggaga
     2761 ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc cggagactgc gagatcatag
     2821 atatagatct cactacgcgg ctgctcaaac ttgggcagaa cgtaagccgc gagagcgcca
     2881 acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg tcttactacg gagcaagttc
     2941 ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg ctacgtctcc gaactcacga
     3001 ccgaaaagat caagagcagc ccgcatggat ttgacttggt cagggccgag cctacatgtg
     3061 cgaatgatgc ccatacttga gccacctaac tttgttttag ggcgactgcc ctgctgcgta
     3121 acatcgttgc tgctgcgtaa catcgttgct gctccataac atcaaacatc gacccacggc
     3181 gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac agtcataaca
     3241 agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg ttctggacca
     3301 gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg cttatgtcaa
     3361 ctgggtttgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct tgggcagcag
     3421 cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg tctccacgca
     3481 tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca cggatctgcc
     3541 cttgcttcag gagatcggta gacctcggcc gtcgcggcgc ttgccggtgg tgctgacccc
     3601 ggatgaagtg gttcgcatcc tcggttttct ggaaggcgag catcgtttgt tcgcccagga
     3661 ctctagctat agttctagtg gttggctaca gctttgtttg tactatcaac aggttgaact
     3721 gctgatcaac agatcctcta cgcggccgcg gtaccataac ttcgtatagc atacattata
     3781 cgaagttatc tggcctaggt taattaaacg ctctatggtc taaagtttaa acctaatgat
     3841 gcctgatgtt tcaatcagga gatccgaacc agataagtga aatctagttc caaactattt
     3901 tgtcattttt aattttcgta ttagcttacg acgctacacc cagttcccat ctattttgtc
     3961 actcttccct aaataatcct taaaaactcc atttccaccc ctcccagttc ccaactattt
     4021 tgtccgccca cagcggggca tttttcttcc tgttatgttt ttaatcaaac atcctgccaa
     4081 ctccatgtga caaaccgtca tcttcggcta cttt
//
