LOCUS       sequence-442572-        2925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and pBR322 origin
            of replication. Carries E-F 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_EF
FEATURES             Location/Qualifiers
     source          1..2925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     292..311
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(378..480)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(524..940)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     531..549
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(553..596)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    599..620
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     835..857
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     849..866
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     850..866
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    867..923
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(936..952)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(936..952)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(949..971)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    960..976
                     /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(984..990,991..1008,1009..1014))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1029..1050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1361..1552
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1546..1568)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1654..1793
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1709..1728
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1808..1825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1979..2567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2059..2078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2598..2637)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(2643..2925,1..377))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
       61 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
      121 tcccatatca ccagctcacc gtctttcatt gccatacgta attccggatg agcattcatc
      181 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
      241 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
      301 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
      361 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgacaa ctcaaaaaat
      421 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgccgatca
      481 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
      541 tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga
      601 attcgcggcc gcttctagag actagtgtgg aagacatgct tagagacctg caccatatgc
      661 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
      721 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
      781 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
      841 cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacccgggga tcctctagag
      901 tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
      961 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1021 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     1081 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg ggtctctcgc
     1141 tatgtcttct aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt
     1201 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc
     1261 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca
     1321 ttaccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     1381 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     1441 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     1501 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     1561 gctcccggcg acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     1621 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     1681 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     1741 cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
     1801 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1861 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1921 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1981 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2041 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2101 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2161 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2221 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2281 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2341 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2401 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2461 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2521 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2581 tcttttctac cggtccgtcc aaaaaaaaag gctccaaaag gagcctttaa ttgtatcggt
     2641 gattacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     2701 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     2761 tcgccttgcg tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg
     2821 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgatac gaaaaacata
     2881 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaa
//
