LOCUS       sequence-405404-        2717 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Receiver cloning vector for odd-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 2717)
  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 2717)
  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-4
FEATURES             Location/Qualifiers
     source          1..2717
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      801..1389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1290..1309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1647..1970)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1652..1671
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1865..1887
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1879..1896
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1880..1896
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1897..1953
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1966..1982)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1966..1982)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1979..2001)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1990..2006
                     /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(2014..2020,2021..2038,2039..2044))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2059..2080
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             join(2615..2717,1..713)
                     /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             join(2615..2717,1..713)
                     /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(2688..2707)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aatcaggtgc gacaatctac cgattgtatg ggaagcccga tgcgccagag ttgtttctga
       61 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
      121 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
      181 ggttactcac cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg
      241 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
      301 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg actcgctcag gcgcaatcac
      361 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
      421 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
      481 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
      541 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
      601 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
      661 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcaccta
      721 gaatgcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
      781 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
      841 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
      901 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
      961 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1021 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1081 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1141 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1201 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1261 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1321 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1381 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1441 gctcacatgt tctttcctgc gttatcccct gattctgtgg gtggaagggc tcggagttgt
     1501 ggtaatctat gtatcctggg gtgctcttca acctagcgtg agaccgtgct cactagtgtc
     1561 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1621 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     1681 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     1741 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1801 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1861 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tcgagctcgg tacccgggga
     1921 tcctctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
     1981 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     2041 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     2101 cgctttccag tcgggaaacc tgtcgtgcca gctgcgctgt caatggtctc actcctctgt
     2161 gaagagctcg gagacgagac gagacagcct gagaatggat gcgagtaatg agctcggcac
     2221 aaaatcacca ctcgatacag gcagcccatc agtccgggac ggcgtcagcg ggagagccgt
     2281 tgtaaggcgg cagactttgc tcatgttacc gatgctattc ggaagaacgg caactaagct
     2341 gccgggtttg aaacacggat gatctcgcgg agggtagcat gttgattgta acgatgacag
     2401 agcgttgctg cctgtgatca ccgcggtttc aaaatcggct ccgtcgatac tatgttatac
     2461 gccaactttg aaaacaactt tgaaaaagct gttttctggt atttaaggtt ttagaatgca
     2521 aggaacagtg aattggagtt cgtcttgtta taattagctt cttggggtat ctttaaatac
     2581 tgtagaaaag aggaaggaaa taataagttc tgttatgagc catattcaac gggaaacgtc
     2641 ttgctcaagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
     2701 tcgcgataat gtcgggc
//
