LOCUS       sequence-405418-        2686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Receiver cloning vector for even-4 level constructs. Contains lacZ 
            for blue/white screening..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2686)
  AUTHORS   Pollak B, Matute T, Nunez I, Cerda A, Lopez C, Vargas V, Kan A, 
            Bielinski V, von Dassow P, Dupont CL, Federici F
  TITLE     Universal loop assembly: open, efficient and cross-kingdom DNA 
            fabrication.
  JOURNAL   Synth Biol (Oxf). 2020;5(1):ysaa001. doi: 10.1093/synbio/ysaa001. 
            Epub 2020 Feb 5.
  PUBMED    32161816
REFERENCE   2  (bases 1 to 2686)
  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: "Synth Biol 
            (Oxf)."; date: "2020"; volume: "5(1):ysaa001. doi"; pages: " 
            10.1093/synbio/ysaa001. Epub 2020 Feb 5"
COMMENT     Sequence Label: pCA_e-4
FEATURES             Location/Qualifiers
     source          1..2686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    653..674
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(689..694,695..712,713..719)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    727..743
                     /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     732..754
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     751..767
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     751..767
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             763..1086
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    780..836
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(837..854)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(837..853)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(846..868)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1062..1081)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(1344..1932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1424..1443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2023..2686,1..128))
                     /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"
ORIGIN
        1 ttcaggccgc catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga
       61 tggcgctcga tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct
      121 tccctcatgc gaaacgatcc tcatcctgtc tcttgatcag atattgatcc cctgcgccat
      181 cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca
      241 gagggcgccc cagctggcaa ttccggttcg cttgctaagc ttgcatgcct gcaagtcgac
      301 tctagaggag cggtgatcac aggcagcaac gctctgtcat cgttacaatc aacatgctac
      361 cctccgcgag atcatccgtg tttcaaaccc ggcagcttag ttgccgttct tccgaatagc
      421 atcggtaaca tgagcaaagt ctgccgcctt acaacggctc tcccgctgac gccgtcccgg
      481 actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctcattact cgcatccatt
      541 ctcaggctgt ctcgtctcgt ctccgaggtc tcagcttaat gtgaagagct tgacaccgca
      601 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      661 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      721 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      781 agcttgcatg cctgcaggtc gactctagag gatccccggg taccgagctc gaattcactg
      841 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
      901 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      961 tcccaacagt tgcgcagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     1021 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     1081 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     1141 ctgctcccgg catccgctta cagacaagct gtgacactag tgagcacgct cttcaggttc
     1201 gcttgagacc accccaggat acatagatta ccacaactcc gagcccttcc acccacagaa
     1261 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1321 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1381 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1441 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1501 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1561 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1621 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1681 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1741 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     1801 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1861 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1921 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1981 aactcacgtt aagggatttt ggtcatgcat tctaggtgat tattatttgc cgactacctt
     2041 ggtgatctcg cctttcacgt agtggacaaa ttcttccaac tgatctgcgc gcgaggccaa
     2101 gcgatcttct tcttgtccaa gataagcctg tctagcttca agtatgacgg gctgatactg
     2161 ggccggcagg cgctccattg cccagtcggc agcgacatcc ttcggcgcga ttttgccggt
     2221 tactgcgctg taccaaatgc gggacaacgt aagcactaca tttcgctcat cgccagccca
     2281 gtcgggcggc gagttccata gcgttaaggt ttcatttagc gcctcaaata gatcctgttc
     2341 aggaaccgga tcaaagagtt cctccgccgc tggacctacc aaggcaacgc tatgttctct
     2401 tgcttttgtc agcaagatag ccagatcaat gtcgatcgtg gctggctcga agataccagc
     2461 aagaatgtca ttgcgctgcc attctccaaa ttgcagttcg cgcttagctg gataacgcca
     2521 cggaatgatg tcgtcgtgca caacaatggt gacttctaca gcgcggagaa tctcactctc
     2581 tccaggggaa gccgaagttt ccaaaaggtc gttgatcaaa gctcgccgcg ttgtttcatc
     2641 aagccttacg gtcaccgtaa ccagcaaatc aatatcactg tgtggc
//
