LOCUS       sequence-442571-        2925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and pBR322 origin
            of replication. Carries F-G 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_FG
FEATURES             Location/Qualifiers
     source          1..2925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     227..246
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(313..415)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(459..875)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLAMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     466..484
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(488..531)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    534..555
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     770..792
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     784..801
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     785..801
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    802..858
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(871..887)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(871..887)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(884..906)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    895..911
                     /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(919..925,926..943,944..949))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    964..985
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1296..1487
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1481..1503)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1589..1728
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1644..1663
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1743..1760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1914..2502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1994..2013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2533..2572)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(2578..2925,1..312))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
       61 tatcaccagc tcaccgtctt tcattgccat acgtaattcc ggatgagcat tcatcaggcg
      121 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
      181 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
      241 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
      301 ttttttctcc attttagctt ccttagctcc tgaaaatctc gacaactcaa aaaatacgcc
      361 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcatgcca
      421 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
      481 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
      541 cggccgcttc tagagactag tgtggaagac atcgctagag acctgcacca tatgcggtgt
      601 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
      661 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
      721 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
      781 cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggatcctc tagagtcgac
      841 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
      901 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
      961 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1021 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcgggggtct cttgccatgt
     1081 cttctaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc
     1141 tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca
     1201 tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc
     1261 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
     1321 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     1381 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     1441 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1501 cggcgacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1561 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     1621 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg
     1681 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     1741 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1801 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1861 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1921 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     1981 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2041 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2101 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2161 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2221 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2281 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2341 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2401 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2461 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2521 tctaccggtc cgtccaaaaa aaaaggctcc aaaaggagcc tttaattgta tcggtgatta
     2581 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2641 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2701 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2761 gtttaaatca aaactggtga aactcaccca gggattggct gatacgaaaa acatattctc
     2821 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     2881 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagag
//
