LOCUS       sequence-442568-        2542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and p15A origin 
            of replication. Carries A-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 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_p15A_AF
FEATURES             Location/Qualifiers
     source          1..2542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..60)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(44..60)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(57..79)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    68..84
                     /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(92..98,99..116,117..122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    137..158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      291..362
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      378..405
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(448..465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(582..1127)
                     /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(662..681)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1241..1335)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1368..2027)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1942..1961
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2028..2130)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(join(2174..2542,1..48))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     2181..2199
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2203..2246)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2249..2270
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     2485..2507
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2499..2516
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2500..2516
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(2517..2542,1..31)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 ctctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag ctgtttcctg
       61 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      121 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      181 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcggggg
      241 tctctcgcta tgtcttctgg atcggttgtc gagtaaggat ctccaggcat caaataaaac
      301 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
      361 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat aggatcctaa
      421 ctcgagccta gggatatatt ccgcttcctc gctcactgac tcgctacgct cggtcgttcg
      481 actgcggcga gcggaaatgg cttacgaacg gggcggagat ttcctggaag atgccaggaa
      541 gatacttaac agggaagtga gagggccgcg gcaaagccgt ttttccatag gctccgcccc
      601 cctgacaagc atcacgaaat ctgacgctca aatcagtggt ggcgaaaccc gacaggacta
      661 taaagatacc aggcgtttcc ccctggcggc tccctcgtgc gctctcctgt tcctgccttt
      721 cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt ctcattccac gcctgacact
      781 cagttccggg taggcagttc gctccaagct ggactgtatg cacgaacccc ccgttcagtc
      841 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggaaa gacatgcaaa
      901 agcaccactg gcagcagcca ctggtaattg atttagagga gttagtcttg aagtcatgcg
      961 ccggttaagg ctaaactgaa aggacaagtt ttggtgactg cgctcctcca agccagttac
     1021 ctcggttcaa agagttggta gctcagagaa ccttcgaaaa accgccctgc aaggcggttt
     1081 tttcgttttc agagcaagag attacgcgca gaccaaaacg atctcaagaa gatcatctta
     1141 ttaatcagat aaaatatttc tagatttcag tgcaatttat ctcttcaaat gtagcacctg
     1201 aagtcagccc catacgatat aagttgttac tagtgcttgg attctcacca ataaaaaacg
     1261 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     1321 ctatcaacag gagtccaagc gagctctcga accccagagt cccgcgatta cgccccgccc
     1381 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     1441 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     1501 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     1561 aaactggtga aactcaccca gggattggct gatacgaaaa acatattctc aataaaccct
     1621 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     1681 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     1741 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     1801 gccatacgta attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     1861 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     1921 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     1981 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     2041 gctcctgaaa atctcgacaa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2101 tgaaagttgg aacctcttac gtgccgatca tgccacctga cgtctaagaa accattatta
     2161 tcatgacatt aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat
     2221 ccttagcttt cgctaaggat gatttctgga attcgcggcc gcttctagag actagtgtgg
     2281 aagacatgga gagagacctg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     2341 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     2401 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     2461 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat
     2521 tcgagctcgg tacccgggga tc
//
