LOCUS       sequence-442567-        2542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and p15A 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 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_FG
FEATURES             Location/Qualifiers
     source          1..2542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..37
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     29..46
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     30..46
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    47..103
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(116..132)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(116..132)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(129..151)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    140..156
                     /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(164..170,171..188,189..194))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    209..230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      363..434
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      450..477
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(520..537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(654..1199)
                     /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(734..753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1313..1407)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1440..2099)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2014..2033
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2100..2202)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(join(2246..2542,1..120))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLAMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     2253..2271
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2275..2318)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2321..2342
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgagctcgg
       61 tacccgggga tcctctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat
      121 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
      181 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
      241 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
      301 aacgcgcggg ggtctcttgc catgtcttct ggatcggttg tcgagtaagg atctccaggc
      361 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
      421 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
      481 ataggatcct aactcgagcc tagggatata ttccgcttcc tcgctcactg actcgctacg
      541 ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag atttcctgga
      601 agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc gtttttccat
      661 aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg gtggcgaaac
      721 ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt gcgctctcct
      781 gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt gtctcattcc
      841 acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta tgcacgaacc
      901 ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccgga
      961 aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag gagttagtct
     1021 tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac tgcgctcctc
     1081 caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa aaaccgccct
     1141 gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa cgatctcaag
     1201 aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt atctcttcaa
     1261 atgtagcacc tgaagtcagc cccatacgat ataagttgtt actagtgctt ggattctcac
     1321 caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg
     1381 tcattactgg atctatcaac aggagtccaa gcgagctctc gaaccccaga gtcccgcgat
     1441 tacgccccgc cctgccactc atcgcagtac tgttgtaatt cattaagcat tctgccgaca
     1501 tggaagccat cacaaacggc atgatgaacc tgaatcgcca gcggcatcag caccttgtcg
     1561 ccttgcgtat aatatttgcc catggtgaaa acgggggcga agaagttgtc catattggcc
     1621 acgtttaaat caaaactggt gaaactcacc cagggattgg ctgatacgaa aaacatattc
     1681 tcaataaacc ctttagggaa ataggccagg ttttcaccgt aacacgccac atcttgcgaa
     1741 tatatgtgta gaaactgccg gaaatcgtcg tggtattcac tccagagcga tgaaaacgtt
     1801 tcagtttgct catggaaaac ggtgtaacaa gggtgaacac tatcccatat caccagctca
     1861 ccgtctttca ttgccatacg taattccgga tgagcattca tcaggcgggc aagaatgtga
     1921 ataaaggccg gataaaactt gtgcttattt ttctttacgg tctttaaaaa ggccgtaata
     1981 tccagctgaa cggtctggtt ataggtacat tgagcaactg actgaaatgc ctcaaaatgt
     2041 tctttacgat gccattggga tatatcaacg gtggtatatc cagtgatttt tttctccatt
     2101 ttagcttcct tagctcctga aaatctcgac aactcaaaaa atacgcccgg tagtgatctt
     2161 atttcattat ggtgaaagtt ggaacctctt acgtgccgat catgccacct gacgtctaag
     2221 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca
     2281 gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg ccgcttctag
     2341 agactagtgt ggaagacatc gctagagacc tgcaccatat gcggtgtgaa ataccgcaca
     2401 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     2461 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     2521 ctgcaaggcg attaagttgg gt
//
