LOCUS       sequence-442566-        2542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and p15A origin 
            of replication. Carries G-H 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_GH
FEATURES             Location/Qualifiers
     source          1..2542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     297..316
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(383..485)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(529..945)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLAMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     536..554
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(558..601)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    604..625
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     840..862
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     854..871
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     855..871
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    872..928
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(941..957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(941..957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(954..976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    965..981
                     /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(989..995,996..1013,1014..1019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1034..1055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1188..1259
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1275..1302
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1345..1362)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1479..2024)
                     /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(1559..1578)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2138..2232)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2265..2542,1..382))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
       61 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
      121 cactatccca tatcaccagc tcaccgtctt tcattgccat acgtaattcc ggatgagcat
      181 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
      241 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
      301 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
      361 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gacaactcaa
      421 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
      481 gatcatgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
      541 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
      601 ctggaattcg cggccgcttc tagagactag tgtggaagac attgccagag acctgcacca
      661 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      721 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      781 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      841 ccagtcacga cgttgtaaaa cgacggccag tgaattcgag ctcggtaccc ggggatcctc
      901 tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
      961 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1021 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1081 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcgggggtct
     1141 ctactaatgt cttctggatc ggttgtcgag taaggatctc caggcatcaa ataaaacgaa
     1201 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
     1261 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagg atcctaactc
     1321 gagcctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     1381 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
     1441 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
     1501 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     1561 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     1621 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     1681 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     1741 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     1801 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     1861 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     1921 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     1981 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     2041 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     2101 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc
     2161 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     2221 tcaacaggag tccaagcgag ctctcgaacc ccagagtccc gcgattacgc cccgccctgc
     2281 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
     2341 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
     2401 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
     2461 ctggtgaaac tcacccaggg attggctgat acgaaaaaca tattctcaat aaacccttta
     2521 gggaaatagg ccaggttttc ac
//
