LOCUS       sequence-369475-        6645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 golden gate cloning vector for low copy number stable 
            environmental gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6645)
  AUTHORS   Geddes BA, Mendoza-Suarez MA, Poole PS
  TITLE     A Bacterial Expression Vector Archive (BEVA) for Flexible Modular 
            Assembly of Golden Gate-Compatible Vectors.
  JOURNAL   Front Microbiol. 2019 Jan 14;9:3345. doi: 10.3389/fmicb.2018.03345. 
            eCollection 2018.
  PUBMED    30692983
REFERENCE   2  (bases 1 to 6645)
  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: "Front 
            Microbiol."; date: "2019-01-14"; volume: "9:3345. doi"; pages: " 
            10.3389/fmicb.2018.03345. eCollection 2018"
COMMENT     Sequence Label: pOGG026
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..324
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    18..74
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(75..92)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(75..91)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(84..106)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(300..319)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      429..523
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(622..641)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             633..1448
                     /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"
     oriT            1599..1707
                     /note="incP origin of transfer"
     CDS             complement(1768..2916)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      3309..3840
                     /label=oriV
                     /note="incP origin of replication"
     terminator      complement(6362..6448)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    6536..6557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6572..6577,6578..6595,6596..6602)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6610..6626
                     /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)."
     primer_bind     6615..6637
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6634..6645,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6634..6645,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta
       61 ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
      121 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
      181 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
      241 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
      301 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
      361 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gacggtctca
      421 cgctcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
      481 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaggca attgtgtctc
      541 aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt
      601 ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcagcgt gaaacgagct
      661 gtagccgtcc gcgtctgaac agcaacatgg atgcggatct gtatggctat aaatgggcgc
      721 gtgataacgt gggtcagagc ggcgcgacca tttatcgtct gtatggcaaa ccggatgcgc
      781 cggaactgtt tctgaaacat ggcaaaggca gcgtggcgaa cgatgtgacc gatgaaatgg
      841 tgcgtctgaa ctggctgacc gaatttatgc cgctgccgac cattaaacat tttattcgca
      901 ccccggatga tgcgtggctg ctgaccaccg cgattccggg caaaaccgcg tttcaggtgc
      961 tggaagaata tccggatagc ggcgaaaaca ttgtggatgc gctggccgtg tttctgcgtc
     1021 gtctgcatag cattccggtg tgcaactgcc cgtttaacag cgatcgtgtg tttcgtctgg
     1081 cccaggcgca gagccgtatg aacaacggcc tggtggatgc gagcgatttt gatgatgaac
     1141 gtaacggctg gccggtggaa caggtgtgga aagaaatgca taaactgctg ccgtttagcc
     1201 cggatagcgt ggtgacccac ggcgatttta gcctggataa cctgattttc gatgaaggca
     1261 aactgattgg ctgcattgat gtgggccgtg tgggcattgc ggatcgttat caggatctgg
     1321 ccattctgtg gaactgcctg ggcgaattta gcccgagcct gcaaaaacgt ctgtttcaga
     1381 aatatggcat tgataatccg gatatgaaca aactgcaatt tcatctgatg ctggatgaat
     1441 ttttctaata attaattgaa ctacttttcc gctgcataac cctgcttcgg ggtcattata
     1501 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     1561 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     1621 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     1681 gggaatcctg ctctgcgagg ctggccgtag gccggccgcg atgcaggtgg ctgctgaacc
     1741 cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac
     1801 ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc
     1861 gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt
     1921 gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga
     1981 cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac
     2041 gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg
     2101 gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat
     2161 cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat
     2221 cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg
     2281 ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag
     2341 ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg
     2401 cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc
     2461 gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat
     2521 ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggctt cgctgacctg
     2581 ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc
     2641 gatggtcatc gacttcgcca aacctgccgc ctcctgttcg agccgacgcg aacgctccac
     2701 ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt
     2761 ggccgtagct tgctggacta tcgagccgac ggactggaag gtttcgcggg gcgcacgcat
     2821 gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc
     2881 ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc
     2941 ggaattaatt ccccggatcg atccgtcgat cttgatcccc tgcgccatca gatccttggc
     3001 ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga gggcgcccca
     3061 gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta tcgccatgta
     3121 agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc
     3181 cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt ctacgtggct
     3241 gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg
     3301 cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg
     3361 tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc
     3421 aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga
     3481 ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact
     3541 ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg
     3601 tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta
     3661 aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg
     3721 agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc
     3781 cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc
     3841 ctacatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga tcccccgcag
     3901 aaaaaaagga tctcaagaag atcctttgat cttttctaca ttactgcatg agcttgtgga
     3961 agtgtgctga gcataaatga ttctcctagc tgttctttgg gtaagtacgc catcaggacg
     4021 ttgtgagtgg cgcgattttt agcggctgaa atcagccctt gagcctgtcg gcaagtcgcg
     4081 tcatgaggtc catgcgctca tgcaggatcg ccacgaccaa cgcgggttcg cccgcacgcg
     4141 gcaggcaaaa aacgtagtgg tgttcgcagc gggccatccg cagcgcggga aagagttcgc
     4201 tcatgtcctt aaacgggcct tcgccggcgg caagcctggc tatgccctgt tccagcttag
     4261 cgatatagcg gcgcacctgc gccgcgcccc actcccggcg cgtgtagcgg atgatgccgc
     4321 gtagatcggc ttcggcctca gccgtgagga tgtaggccgt caagcgcgat ccccgctgag
     4381 ttcttcatca agaatttcgc cgacgctctt ggtggacacc ttgccggcaa gcccatcgtt
     4441 gatgcggttc cccagcatgg ttttcagttc ctgccatgcc tgatcggcat cagcgtcacc
     4501 ggggaacaga cgttcgaggg cgtattgctt aatggtcttg ccctgcaagg cggccagggc
     4561 tttcaggctc tggtgctgct ggtccgtcat gtcgattgtc aggcggctca ttggataacc
     4621 tccataaaat acacgtaacc acattagcac atatgtgggc gtgaggctac agcgcgaggc
     4681 gcattaaggt cgggaaaatg cgctaggcgc atttaaattg cgtattgctg taatgcgcca
     4741 tgccggctag actaggccca aatgggtata cccaatttga ccaaggggga cgcgatgagg
     4801 gcggccaagc actaccgaca acttctatcc atcgacttca acatcgaggc gctggccttc
     4861 gtgcctggac ccgacggcac acgcggccgg cgcatccacg tcctggggcg cgaggtccgc
     4921 gaccggcccg gcctggtcga gtacctttcg ccggcgttcg gctcgcgggt ggcgctggac
     4981 ggctactgca aggccaattt cgatgcagtg ctgcacctgg cgtaccccga tcatcagcaa
     5041 tggggccacg catgaagcgc cgaagctacg ccatgctgcg cgccgctgcc gcgctggccg
     5101 tcctggtcgt tgcctcgccg gcatgggccg agctgcgcgg cgaggtcgtg cgcatcatcg
     5161 acggcgacac catcgacgtg ctggtagaca agcagccggt gcgcgtgcgc ctggtggaca
     5221 ttgacgcgcc ggaaaagcgg caagccttcg gcgaacgtgc gcgccaggcg ctggccggca
     5281 tggtgttccg ccggcacgtc ctggtcgacg agaaggacac cgaccgttac ggccgcacgc
     5341 tgggcaccgt ctgggtcaac atggagctgg ccagccggcc gccgcagccg cgcaacgtca
     5401 acgccgcgat ggttcaccag ggcatggcgt gggcctatcg cttccacggc cgcgcggccg
     5461 accctgaaat gctgcggctc gaacaggagg cgcgaggcaa gcgcgtcggc ctctggtccg
     5521 atccgcacgc cgtcgagccg tggaaatggc gacgcgagag caacaaccgg agggacgaag
     5581 gttgaaggtc gcccgcatct acctgcgcgc cagtacggac gagcagaatc ttgaacgcca
     5641 ggagagcctt gtagcggcca cgcgggccgc cgggtactac gtcgccggca tctaccgcga
     5701 gaaggcgtcc ggcgcacgcg ccgaccggcc cgagctgctg cgcatgatcg cggacctgca
     5761 acctggtgaa gtcgtcgttg cggagaagat cgaccgcatc agccgcttgc cgttggccga
     5821 ggccgagcgc ctggttgcgt cgatccgggc caaaggggcc aagctggccg tgcctggcgt
     5881 ggtggacctg tcggagctgg ccgccgaggc gaacggagtg gcgaaaatcg ttctggaatc
     5941 cgtccaggac atgcttttga agctcgcctt gcagatggcc cgcgacgact acgaggatcg
     6001 gcgcgagcgt caacgtcagg gtgtccagtt ggcgaaggcc gccggccgct acaccggccg
     6061 caaacgtgac gccggcatgc acgaccgcat catcacgctt cgctccggcg gatcgagcat
     6121 tgccaagacg gccaagctgg tcggatgcag cccgagccag gtcaaacgag tgtgggcggc
     6181 ctggaacgcg cagcagcaaa aataaagccg ggcagtgccc ggcttttctc accttttcgc
     6241 gtcccgcagg gccgctgcga gcgccctacc tagatcctcg ctttccccct cggtgtagtc
     6301 cggccagggc acgaagggcg cggatgcgaa cctgttgagc aggtcagatc tagggcggcg
     6361 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
     6421 ctttcgactg agcctttcgt tttatttgat gcctgccgaa ttcggatccg gagtgagacc
     6481 gcagctggca cgacaggttt gccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6541 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6601 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagct
//
