LOCUS       sequence-468600-        2542 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with chloramphenicol resistance and p15A origin 
            of replication. Carries A-E 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_AE
FEATURES             Location/Qualifiers
     source          1..2542
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     568..587
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(654..756)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             complement(800..1216)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TLVSRSGREFQKSSLAKAKDFFYLKFCLVIRLFL"
     primer_bind     807..825
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(829..872)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    875..896
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     1111..1133
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1125..1142
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1126..1142
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1143..1199
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1212..1228)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1212..1228)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1225..1247)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1236..1252
                     /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(1260..1266,1267..1284,1285..1290))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1305..1326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1459..1530
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1546..1573
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1616..1633)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1750..2295)
                     /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(1830..1849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2409..2503)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2536..2542,1..653))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
       61 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
      121 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
      181 aaatcaaaac tggtgaaact cacccaggga ttggctgata cgaaaaacat attctcaata
      241 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
      301 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
      361 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
      421 ttcattgcca tacgtaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
      481 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
      541 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
      601 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
      661 tccttagctc ctgaaaatct cgacaactca aaaaatacgc ccggtagtga tcttatttca
      721 ttatggtgaa agttggaacc tcttacgtgc cgatcatgcc acctgacgtc taagaaacca
      781 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa
      841 aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt ctagagacta
      901 gtgtggaaga catggagaga gacctgcacc atatgcggtg tgaaataccg cacagatgcg
      961 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
     1021 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     1081 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
     1141 gtgaattcga gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg
     1201 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1261 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1321 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1381 cattaatgaa tcggccaacg cgcgggggtc tctgcttatg tcttctggat cggttgtcga
     1441 gtaaggatct ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1501 tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt
     1561 gggcctttct gcgtttatag gatcctaact cgagcctagg gatatattcc gcttcctcgc
     1621 tcactgactc gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg
     1681 gcggagattt cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc
     1741 aaagccgttt ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa
     1801 tcagtggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggcggctc
     1861 cctcgtgcgc tctcctgttc ctgcctttcg gtttaccggt gtcattccgc tgttatggcc
     1921 gcgtttgtct cattccacgc ctgacactca gttccgggta ggcagttcgc tccaagctgg
     1981 actgtatgca cgaacccccc gttcagtccg accgctgcgc cttatccggt aactatcgtc
     2041 ttgagtccaa cccggaaaga catgcaaaag caccactggc agcagccact ggtaattgat
     2101 ttagaggagt tagtcttgaa gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt
     2161 ggtgactgcg ctcctccaag ccagttacct cggttcaaag agttggtagc tcagagaacc
     2221 ttcgaaaaac cgccctgcaa ggcggttttt tcgttttcag agcaagagat tacgcgcaga
     2281 ccaaaacgat ctcaagaaga tcatcttatt aatcagataa aatatttcta gatttcagtg
     2341 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgttacta
     2401 gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc
     2461 agatggagtt ctgaggtcat tactggatct atcaacagga gtccaagcga gctctcgaac
     2521 cccagagtcc cgcgattacg cc
//
