LOCUS       sequence-72955-e        2671 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Destination vector for Golden Gate assembly; Few restriction sites 
            for subcloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2671)
  AUTHORS   Kirchmaier S, Lust K, Wittbrodt J
  TITLE     Golden GATEway cloning--a combinatorial approach to generate fusion 
            and recombination constructs.
  JOURNAL   PLoS One. 2013 Oct 7;8(10):e76117. doi: 
            10.1371/journal.pone.0076117.
  PUBMED    24116091
REFERENCE   2  (bases 1 to 2671)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2013-10-7"; volume: "8(10):e76117. doi"; pages: " 
            10.1371/journal.pone.0076117"
COMMENT     Sequence Label: pGGDestSC+ATG
FEATURES             Location/Qualifiers
     source          1..2671
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      300..888
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     789..808
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1154..1163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1176..1559)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1325..1347
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1339..1356
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1340..1356
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1363..1381
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1412..1421
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1416..1425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1519..1537)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1520..1537)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1555..1571)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1568..1590)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1579..1595
                     /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(1603..1609,1610..1627,1628..1633))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1648..1669
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     1825..1841
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1826..1842
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1863..1881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1886..1902)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2004..2023)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(2015..2671,1..153)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2082..2101)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
       61 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
      121 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
      181 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
      241 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
      301 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
      361 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
      421 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
      481 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
      541 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
      601 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
      661 tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg
      721 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
      781 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
      841 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
      901 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
      961 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     1021 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     1081 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagtatagg cgcgccaggg
     1141 cgaattgggt accgccgcca tggcgagacc cgcgcttaat gcgccgctac agggcgcgtc
     1201 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1261 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1321 ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga ctcactatag
     1381 ggcgaattgg gcccgacgtc gcatgctccc ggccgccatg gcggccgcgg gaattcgata
     1441 tcactagtga attcgcggcc gcctgcaggt cgaccatatg ggagagctcc caacgcgttg
     1501 gatgcatagc ttgagtattc tatagtgtca cctaaatagc ttggcgtaat catggtcata
     1561 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1621 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1741 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     1801 gctgcgctcg gggtctctta aggctcgagg tcgacggtat cggccggcct aatctgatat
     1861 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1921 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1981 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     2041 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     2101 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     2161 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     2221 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     2281 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     2341 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     2401 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     2461 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     2521 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     2581 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     2641 aataggttgt attgatgttg gacgagtcgg a
//
