LOCUS       sequence-72954-e        2665 bp    DNA     circular SYN 15-DEC-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 2665)
  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 2665)
  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: "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..2665
                     /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"
     CDS             complement(1170..1553)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSYLGDTIEYSSYASNALGALPYGRPAGGREFTSDIEFPR
                     PPWRPGACDVGPNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEA
                     RTDRPSQQLRSLNGEWTRPVAAH"
     primer_bind     1319..1341
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1333..1350
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1334..1350
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1357..1375
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1406..1415
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1410..1419
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(1513..1531)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1514..1531)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1549..1565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1562..1584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1573..1589
                     /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(1597..1603,1604..1621,1622..1627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1642..1663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     1819..1835
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1820..1836
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(1857..1875)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1880..1896)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1998..2017)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(2009..2665,1..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(2076..2095)
                     /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 accgtatcga gacccgcgct taatgcgccg ctacagggcg cgtccattcg
     1201 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     1261 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     1321 cagtcacgac gttgtaaaac gacggccagt gaattgtaat acgactcact atagggcgaa
     1381 ttgggcccga cgtcgcatgc tcccggccgc catggcggcc gcgggaattc gatatcacta
     1441 gtgaattcgc ggccgcctgc aggtcgacca tatgggagag ctcccaacgc gttggatgca
     1501 tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt catagctgtt
     1561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1801 ctcggggtct cttaaggctc gaggtcgacg gtatcggccg gcctaatctg atatccccta
     1861 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     1921 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1981 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     2041 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     2101 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     2161 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2221 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2281 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2341 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2401 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2461 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2521 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2581 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2641 ttgtattgat gttggacgag tcgga
//
