LOCUS       sequence-405416-        2686 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Receiver cloning vector for even-2 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-2
FEATURES             Location/Qualifiers
     source          1..2686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      765..1353
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1254..1273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1611..1934)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     primer_bind     1616..1635
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1829..1851
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1843..1860
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1844..1860
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1861..1917
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1930..1946)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1930..1946)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1943..1965)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1954..1970
                     /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(1978..1984,1985..2002,2003..2008))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2023..2044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             join(2569..2686,1..674)
                     /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 gcggcctgaa gccacacagt gatattgatt tgctggttac ggtgaccgta aggcttgatg
       61 aaacaacgcg gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga
      121 gtgagattct ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc
      181 gttatccagc taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgctg
      241 gtatcttcga gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag
      301 aacatagcgt tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac
      361 aggatctatt tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg
      421 ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg
      481 gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt
      541 atcagcccgt catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg
      601 cctcgcgcgc agatcagttg gaagaatttg tccactacgt gaaaggcgag atcaccaagg
      661 tagtcggcaa ataataatca cctagaatgc atgaccaaaa tcccttaacg tgagttttcg
      721 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
      781 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
      841 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
      901 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
      961 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1021 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1081 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1141 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1201 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1261 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1321 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1381 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1441 gtgggtggaa gggctcggag ttgtggtaat ctatgtatcc tggggtggtc tcacatttac
     1501 ctgaagagcg tgctcactag tgtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1561 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     1621 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     1681 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     1741 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     1801 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     1861 gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc
     1921 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1981 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     2041 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgcg
     2101 ctgtcaagct cttcacatta gtatgagacc tcggagacga gacgagacag cctgagaatg
     2161 gatgcgagta atgagctcgg cacaaaatca ccactcgata caggcagccc atcagtccgg
     2221 gacggcgtca gcgggagagc cgttgtaagg cggcagactt tgctcatgtt accgatgcta
     2281 ttcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag
     2341 catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcaccgctcc tctagagtcg
     2401 acttgcaggc atgcaagctt agcaagcgaa ccggaattgc cagctggggc gccctctggt
     2461 aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg
     2521 cgcaggggat caatatctga tcaagagaca ggatgaggat cgtttcgcat gagggaagcg
     2581 gtgatcgccg aagtatcgac tcaactatca gaggtagttg gcgtcatcga gcgccatctc
     2641 gaaccgacgt tgctggccgt acatttgtac ggctccgcag tggatg
//
