LOCUS       sequence-442569-        2925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and pBR322 origin
            of replication. Carries A-E 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_AE
FEATURES             Location/Qualifiers
     source          1..2925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     287..306
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(373..475)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(519..935)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     526..544
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(548..591)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    594..615
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     830..852
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     844..861
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     845..861
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    862..918
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(931..947)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(931..947)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(944..966)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    955..971
                     /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(979..985,986..1003,1004..1009))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1024..1045
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1356..1547
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1541..1563)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1649..1788
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1704..1723
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1803..1820)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1974..2562)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2054..2073)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2593..2632)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(2638..2925,1..372))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
       61 cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
      121 tatcaccagc tcaccgtctt tcattgccat acgtaattcc ggatgagcat tcatcaggcg
      181 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
      241 aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa
      301 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
      361 ttttttctcc attttagctt ccttagctcc tgaaaatctc gacaactcaa aaaatacgcc
      421 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcatgcca
      481 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
      541 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
      601 cggccgcttc tagagactag tgtggaagac atggagagag acctgcacca tatgcggtgt
      661 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
      721 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
      781 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
      841 cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggatcctc tagagtcgac
      901 ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
      961 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     1021 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     1081 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcgggggtct ctgcttatgt
     1141 cttctaacac ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc
     1201 tggtcccgcc gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca
     1261 tgttcatcat cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc
     1321 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
     1381 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     1441 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     1501 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     1561 cggcgacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     1621 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     1681 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg
     1741 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     1801 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1861 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     1921 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     1981 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2041 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2101 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2161 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2221 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2281 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2341 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2401 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2461 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2521 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2581 tctaccggtc cgtccaaaaa aaaaggctcc aaaaggagcc tttaattgta tcggtgatta
     2641 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     2701 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     2761 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
     2821 gtttaaatca aaactggtga aactcaccca gggattggct gatacgaaaa acatattctc
     2881 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgc
//
