LOCUS       sequence-444821-        2542 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Destination vector with chloramphenicol resistance and p15A 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 2542)
  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 2542)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_p15A_EF
FEATURES             Location/Qualifiers
     source          1..2542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..660)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     575..594
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(661..763)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     814..832
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(836..879)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    882..903
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     misc_feature    1150..1206
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1219..1235)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1219..1235)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1232..1254)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1243..1259
                     /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(1267..1273,1274..1291,1292..1297))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1312..1333
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1466..1537
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1553..1580
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1623..1640)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1757..2302)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1837..1856)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2416..2510)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
       61 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
      121 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
      181 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgatacga aaaacatatt
      241 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
      301 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
      361 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
      421 accgtctttc attgccatac gtaattccgg atgagcattc atcaggcggg caagaatgtg
      481 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
      541 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
      601 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
      661 tttagcttcc ttagctcctg aaaatctcga caactcaaaa aatacgcccg gtagtgatct
      721 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcatgccacc tgacgtctaa
      781 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gcagaatttc
      841 agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta
      901 gagactagtg tggaagacat gcttagagac ctgcaccata tgcggtgtga aataccgcac
      961 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt
     1021 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggtgaa agggggatgt
     1081 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaatg
     1141 acggccagtg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc
     1201 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1261 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1321 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1381 ccagctgcat taatgaatcg gccaacgcgc gggggtctct cgctatgtct tctggatcgg
     1441 ttgtcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     1501 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     1561 ttcgggtggg cctttctgcg tttataggat cctaactcga gcctagggat atattccgct
     1621 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     1681 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     1741 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     1801 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1861 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     1921 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     1981 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     2041 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     2101 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     2161 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     2221 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     2281 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat
     2341 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     2401 gttactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa
     2461 caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct
     2521 ctcgaacccc agagtcccgc ga
//
