LOCUS       sequence-402519-        2856 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 RBS acceptor vector with LacZ compatible blue/white 
            screening..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2856)
  TITLE     McCormick Lab Plasmids 2023
REFERENCE   2  (bases 1 to 2856)
  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"
COMMENT     Sequence Label: pCA0.325
FEATURES             Location/Qualifiers
     source          1..2856
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     260..277
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    425..446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(461..466,467..484,485..491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    499..515
                     /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)."
     primer_bind     504..526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     523..539
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     523..539
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             535..858
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    552..608
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(609..626)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(609..625)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(618..640)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(834..853)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(999..1017)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1085..1189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1412..2203
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(2374..2856,1..106)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
       61 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      121 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
      181 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
      241 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
      301 caaaccgcct ctccccgcgc gttggccgat tcattaatca ctctgtggtc tcatactgag
      361 tcttcgtacg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
      421 caattaatgt gagttagctc actcattagg caccctaggc tttacacttt atgcttccgg
      481 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
      541 atgattacgc caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc
      601 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      661 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      721 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
      781 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
      841 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
      901 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgacgta cgaagactca
      961 atgagagacc acgaagtggc tcttcagtgg acgaaagggc ctcgtgatac gcctattttt
     1021 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgcgctcacg
     1201 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     1261 ggcttgttat gactgntttt tttttggggt acagtctatg cctcgggcat ccaagcagca
     1321 agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg ttacgcagca
     1381 gggcagtcgc cctaaaacaa agttaaacat catgagggaa gcggtgatcg ccgaagtatc
     1441 gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc
     1501 cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagcg atattgattt
     1561 gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga tcaacgacct
     1621 tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag aagtcaccat
     1681 tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg
     1741 agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga tcgacattga
     1801 tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag gtccagcggc
     1861 ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa atgaaacctt
     1921 aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt
     1981 gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga
     2041 ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag ctagacaggc
     2101 ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt
     2161 ccattacgta aaaggcgaga tcaccaaggt agtcggcaaa taactgtcag accaagttta
     2221 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     2281 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     2341 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     2401 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     2461 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     2521 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     2581 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     2641 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     2701 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     2761 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     2821 cggcagggtc ggaacaggag agcgcacgag ggagct
//
