LOCUS       sequence-445914-        3427 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4G-cloning donor for mammalian promoter and terminator, C-terminal 
            tag added.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3427)
  AUTHORS   Taschner M, Dickinson JB, Roisne-Hamelin F, Gruber S
  TITLE     4G cloning: rapid gene assembly for expression of multisubunit 
            protein complexes in diverse hosts
  JOURNAL   bioRxiv 2024.06.17.599261
REFERENCE   2  (bases 1 to 3427)
  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: "bioRxiv 
            2024.06.17.599261"
COMMENT     Sequence Label: pD(P+T)_CMVC
FEATURES             Location/Qualifiers
     source          1..3427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        174..276
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             277..936
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(343..362)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1331..1687
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(1740..1773)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        2018..2397
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2398..2601
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2552..2572
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2598..2622
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      3112..3121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3174..3398
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gagacccact gcttgagcct agaagatccg gctgctaaca aagcccgaaa ggaagctgag
       61 ttggctgctg tcaccgctga gcaataacta tcataacccc taggtgccat ttcattacct
      121 ctttctccgc acccgacata gatctgggcc aacttttggc gaaaatgaga cgttgatcgg
      181 cacgtaagag ggtccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
      241 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
      301 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
      361 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
      421 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
      481 gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
      541 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
      601 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
      661 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
      721 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
      781 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
      841 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
      901 caacagtact gcgatgagtg gcagggtggg gcgtaatttt tttaaggcag ttattggtgc
      961 ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat
     1021 tcgaaagcaa attcgacccg gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg
     1081 tctattgctg gtttaccggt ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa
     1141 atgcctgagg ccagtttgct caggctctcc ccgtggaggt aataattgac gatatgatca
     1201 tttattctgc ctcccagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     1261 tctacgtgtt ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg cagcgtgaag
     1321 ctaattccca tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt tgagaggctc
     1381 taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta aaccttaaaa
     1441 gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt agaggctatt
     1501 taagttgctg atttatatta attttattgt tcaaacatga gagcttagta cgtgaaacat
     1561 gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt ttagttcgtt
     1621 aaacatgaga gcttagtacg ttaaacatga gagcttagta cgtgaaacat gagagcttag
     1681 tacgtactat caacaggttg aactgctgat caacagatcc tctacgcggc cgcggtacca
     1741 taacttcgta tagcatacat tatacgaagt tatctgccag gcacatgggt tttactagta
     1801 tcgattcgcg acctactccg gaatattagg tctcagcgac tactccggaa tattaataga
     1861 tcatggactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     1921 ataccgcatc agattggcta ttggccattg catacgttgt atccatatca taatatgtac
     1981 atttatattg gctcatgtcc aacattaccg ccatgttgac attgattatt gactagttat
     2041 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
     2101 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     2161 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     2221 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     2281 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     2341 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
     2401 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
     2461 agtctccacc ccattgacgt caatgggagt ttgttttggc accgaaatca acgggacttt
     2521 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
     2581 gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca
     2641 cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccatcg gctcgcatct
     2701 ctccttcacg cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc
     2761 tgccgcctcc cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc
     2821 tcaggtccag accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc
     2881 tctccacgct ttgcctgacc ctgcttgctc aactctagtt aacggtggag ggcagtgtag
     2941 tctgagcagt actcgttgct gccgcgcgcg ccaccagaca taatagctga cagactaaca
     3001 gactgttcct ttccatgggt cttttctgca gtcaccgtcg tcgacacgtg tgatcagata
     3061 tcatattgaa gagacagagt gatatataaa actgctaact cctaaaaaac cgccaccatg
     3121 gagaccatac gtgtcacggt ctcctaaggt gctttgcaac ggatccagat ctgctgtgcc
     3181 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3241 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3301 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3361 caatagcagg catgctgggg atgcggtggg ctctatggcg ctgagcaata actatcataa
     3421 cctacgg
//
