LOCUS       sequence-461575-        3947 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bigMamAct is evolution of biGBac for mammalian cells. Has empty 
            CMV-driven cassette; users should clone gene of interest into 
            shuttle plasmid prior to multi-assembly step by Gibson cloning .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3947)
  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 3947)
  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: psLIB CMV
FEATURES             Location/Qualifiers
     source          1..3947
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(114..147)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        205..584
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        585..788
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     739..759
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1077..1211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1114..1133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1168..1187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1321..1486
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      1731..2319
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2220..2239
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2473..2490
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2624..2848
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2915..3448)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3637..3642,3643..3659,3660..3665))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tcgtttgttc gcccaggact ctagctatag ttctagtggt tggctacagc tttgtttgta
       61 ctatcaacag gttgaactgc tgatcaacag atcctctacg cggccgcggt accataactt
      121 cgtatagcat acattatacg aagttatctg gttttactag tatcgattcg cgacctactc
      181 cggaatatta atagatcacg cgttgacatt gattattgac tagttattaa tagtaatcaa
      241 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
      301 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
      361 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      421 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      481 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      541 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
      601 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
      661 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
      721 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
      781 gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa atataccgtc
      841 ccaccatcgg gcgcggatcc cggtccgaag cgcgcggaat tcaaaggcct acgtcgacga
      901 gctcactagt cgcggccgct ttcgaatcta gagcctgcag tctcgacaag cttgtcgaga
      961 agtactagag gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     1021 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     1081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     1141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     1201 atgtctggat ctgatcactg cttgagccta gaagatccgg ctgctaacaa agcccgaaag
     1261 gaagctgagt tggctgctgc caccgctgag caataactat cataacccct aggagatccg
     1321 aaccagataa gtgaaatcta gttccaaact attttgtcat ttttaatttt cgtattagct
     1381 tacgacgcta cacccagttc ccatctattt tgtcactctt ccctaaataa tccttaaaaa
     1441 ctccatttcc acccctccca gttcccaact attttgtccg cccacagcgg ggcatttttc
     1501 ttcctgttat gtttttaatc aaacatcctg ccaactccat gtgacaaacc gtcatcttcg
     1561 gctacttttt ctctgtcaca gaatgaaaat ttttctgtca tctcttcgtt attaatgttt
     1621 gtaattgact gaatatcaac gcttatttgc agcctgaatg gcgaatggga cgcgcccgag
     1681 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1741 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     1801 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     1861 cagataccaa atactattct tctagtgtag ccgtagttag gccaccactt caagaactct
     1921 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     1981 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2041 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2101 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2161 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2221 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2281 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2341 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2401 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2461 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     2521 ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa cctggcaaaa
     2581 tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa
     2641 gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac tagacagaat
     2701 agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac ttttgttatg
     2761 gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa agaggggcgt
     2821 ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg aacaactccg
     2881 cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt gggtcgatat
     2941 caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact gcgggatcgt
     3001 caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc tcatgcttga
     3061 ggagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag actgcgagat
     3121 catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa gccgcgagag
     3181 cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta ctacggagca
     3241 agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg tctccgaact
     3301 cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg ccgagcctac
     3361 atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga ctgccctgct
     3421 gcgtaacatc gttgctgctg cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc
     3481 acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa aaaacagtca
     3541 taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt caaggttctg
     3601 gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga acaggcttat
     3661 gtcaactggg tttgtgcctt catccgtttc cacggtgtgc gtcacccggc aaccttgggc
     3721 agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc
     3781 acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct gtgcacggat
     3841 ctgcccttgc ttcaggagat cggtagacct cggccgtcgc ggcgcttgcc ggtggtgctg
     3901 accccggatg aagtggttcg catcctcggt tttctggaag gcgagca
//
