LOCUS       sequence-442570-        2925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and pBR322 origin
            of replication. Carries G-H CIDAR MoClo overhangs for level 1 
            construct cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2925)
  AUTHORS   Nielsen JR, Lewis MJ, Huang WE
  TITLE     Construction and Characterization of MoClo-Compatible Vectors for 
            Modular Protein Expression in E. coli.
  JOURNAL   ACS Synth Biol. 2025 Jan 12. doi: 10.1021/acssynbio.4c00564.
  PUBMED    39801078
REFERENCE   2  (bases 1 to 2925)
  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: "ACS Synth 
            Biol. 2025 Jan 12. doi: 10.1021/acssynbio.4c00564."
COMMENT     Sequence Label: DVC_pBR322_GH
FEATURES             Location/Qualifiers
     source          1..2925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(49..151)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(195..611)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLAMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     202..220
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(224..267)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    270..291
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     506..528
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     520..537
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     521..537
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    538..594
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(607..623)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(607..623)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(620..642)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    631..647
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(655..661,662..679,680..685))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    700..721
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1032..1223
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1217..1239)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1325..1464
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1380..1399
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1479..1496)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1650..2238)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1730..1749)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2269..2308)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(2314..2925,1..48))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2888..2907
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
       61 agctcctgaa aatctcgaca actcaaaaaa tacgcccggt agtgatctta tttcattatg
      121 gtgaaagttg gaacctctta cgtgccgatc atgccacctg acgtctaaga aaccattatt
      181 atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa
      241 tccttagctt tcgctaagga tgatttctgg aattcgcggc cgcttctaga gactagtgtg
      301 gaagacattg ccagagacct gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
      361 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
      421 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
      481 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
      541 ttcgagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggcgta
      601 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
      661 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
      721 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
      781 atgaatcggc caacgcgcgg gggtctctac taatgtcttc taacacctac atctgtatta
      841 acgaagcgct ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc
      901 agttgtttac cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc
      961 gtgagcatcc tctctcgttt catcggtatc attaccccat gaacagaaat cccccttaca
     1021 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     1081 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     1141 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     1201 gtgatgacgg tgaaaacctc tgacacatgc agctcccggc gacggtcaca gcttgtctgt
     1261 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1321 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     1381 ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc
     1441 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1501 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1561 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1621 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1681 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1741 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1801 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1861 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1921 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1981 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2041 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2101 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2161 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2221 agaaaaaaag gatctcaaga agatcctttg atcttttcta ccggtccgtc caaaaaaaaa
     2281 ggctccaaaa ggagccttta attgtatcgg tgattacgcc ccgccctgcc actcatcgca
     2341 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
     2401 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
     2461 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
     2521 cacccaggga ttggctgata cgaaaaacat attctcaata aaccctttag ggaaataggc
     2581 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
     2641 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
     2701 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgtaattc
     2761 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
     2821 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
     2881 acattgagca actgactgaa atgcctcaaa atgttcttta cgatg
//
