LOCUS       sequence-405403-        2722 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Receiver cloning vector for odd-3 level constructs. Contains lacZ 
            for blue/white screening..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2722)
  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 2722)
  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_o-3
FEATURES             Location/Qualifiers
     source          1..2722
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             321..1136
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             321..1136
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(394..413)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1224..1812
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1713..1732
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2070..2393)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     2075..2094
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2288..2310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2302..2319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2303..2319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2320..2376
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2389..2405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2389..2405)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2402..2424)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2413..2429
                     /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(2437..2443,2444..2461,2462..2467))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2482..2503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctttgctcat gttaccgatg ctattcggaa gaacggcaac taagctgccg ggtttgaaac
       61 acggatgatc tcgcggaggg tagcatgttg attgtaacga tgacagagcg ttgctgcctg
      121 tgatcaccgc ggtttcaaaa tcggctccgt cgatactatg ttatacgcca actttgaaaa
      181 caactttgaa aaagctgttt tctggtattt aaggttttag aatgcaagga acagtgaatt
      241 ggagttcgtc ttgttataat tagcttcttg gggtatcttt aaatactgta gaaaagagga
      301 aggaaataat aagttctgtt atgagccata ttcaacggga aacgtcttgc tcaaggccgc
      361 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
      421 ggcaatcagg tgcgacaatc taccgattgt atgggaagcc cgatgcgcca gagttgtttc
      481 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
      541 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
      601 catggttact caccactgcg atcccaggga aaacagcatt ccaggtatta gaagaatatc
      661 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
      721 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgactcgct caggcgcaat
      781 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
      841 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
      901 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
      961 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     1021 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     1081 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcac
     1141 ctagaatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1201 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1261 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1321 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     1381 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1441 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1501 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1561 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1621 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1681 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1741 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1801 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1861 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tgggtggaag ggctcggagt
     1921 tgtggtaatc tatgtatcct ggggtgctct tcactgtagc gtgagaccgt gctcactagt
     1981 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     2041 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     2101 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     2161 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     2221 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     2281 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
     2341 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     2401 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2461 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2521 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcgc tgtcaatggt ctcactcctg
     2581 tatgaagagc tcggagacga gacgagacga gacagcctga gaatggatgc gagtaatgag
     2641 ctcggcacaa aatcaccact cgatacaggc agcccatcag tccgggacgg cgtcagcggg
     2701 agagccgttg taaggcggca ga
//
