LOCUS       sequence-224278-        3417 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 golden gate cloning vector for medium copy number 
            free-living expression .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3417)
  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 3417)
  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: pOGG024
FEATURES             Location/Qualifiers
     source          1..3417
                     /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"
     promoter        join(532..537,538..554,555..560)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             749..1282
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            1476..1584
                     /note="incP origin of transfer"
     CDS             complement(1595..2257)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      2258..3029
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(3134..3220)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     protein_bind    3308..3329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3344..3349,3350..3367,3368..3374)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3382..3398
                     /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     3387..3409
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(3406..3417,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(3406..3417,1..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 attgacataa
      541 gcctgttcgg ttcgtaaact gtaatgcaag tagcgtatgc gctcacgcaa ctggtccaga
      601 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
      661 tgtttttttg tacagcctat gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt
      721 cgatgtttga tgttatggag cagcaacgat gttacgcagc agcaacgatg ttacgcagca
      781 gggcagtcgc cctaaaacaa agttaggtgg ctcaagtatg ggcatcattc gcacatgtag
      841 gctcggccct gaccaagtca aatccatgcg ggctgctctt gatcttttcg gtcgtgagtt
      901 cggcgacgta gccacctact cccaacatca gccggactcc gattacctcg ggaacttgct
      961 ccgtagtaag acattcatcg cgcttgctgc cttcgaccaa gaagcggttg ttggcgctct
     1021 cgcggcttac gttctgccca agtttgagca gccgcgtagt gagatctata tctatgatct
     1081 cgcagtctcc ggagagcacc ggaggcaggg cattgccacc gcgctcatca atctcctcaa
     1141 gcatgaggcc aacgcgcttg gtgcttatgt gatctacgtg caagcagatt acggtgacga
     1201 tcccgcagtg gctctctata caaagttggg catacgggaa gaagtgatgc actttgatat
     1261 cgacccaagt accgccacct aacaattcgt tcaagccgag atcggcttcc cggccgcgga
     1321 gttgttcggt aaattgacta cttttccgct gcataaccct gcttcggggt cattatagcg
     1381 attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt
     1441 gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca
     1501 cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg
     1561 aatcctgctc tgcgaggctg gccgtaggcc ggccctaccg gcgcggcagc gttacccgtg
     1621 tcggcggctc caacggctcg ccatcgtcca gaaaacacgg ctcatcgggc atcggcaggc
     1681 gctgctgccc gcgccgttcc cattcctccg tttcggtcaa ggctggcagg tctggttcca
     1741 tgcccggaat gccgggctgg ctgggcggct cctcgccggg gccggtcggt agttgctgct
     1801 cgcccggata cagggtcggg atgcggcgca ggtcgccatg ccccaacagc gattcgtcct
     1861 ggtcgtcgtg atcaaccacc acggcggcac tgaacaccga caggcgcaac tggtcgcggg
     1921 gctggcccca cgccacgcgg tcattgacca cgtaggccga cacggtgccg gggccgttga
     1981 gcttcacgac ggagatccag cgctcggcca ccaagtcctt gactgcgtat tggaccgtcc
     2041 gcaaagaacg tccgatgagc ttggaaagag tcttttggct gaccaccacg gcgttctggt
     2101 ggcccatctg cgcgacgagg tgatgcagca gcattgccgc cgtgggtttc ctcgcaataa
     2161 gcccggccca cgcctcatgc gctttgcgtt ccgtttggac ccagtgaccg ggcttgttct
     2221 tggcttgaat gccgatttct ctggactgcg tggccatgct tatctccatg cggtaggggt
     2281 gccgcacggt tgcggcacca tgcgcaatca gctgcaactt ttcggcagcg cgacaacaat
     2341 tatgcgttgc gtaaaagtgg cagtcaatta cagattttct ttaacctacg caatgagcta
     2401 ttgcgggggg tgccgcaatg agctgttgcg tacccccctt ttttaagttg ttgattttta
     2461 agtctttcgc atttcgccct atatctagtt ctttggtgcc caaagaaggg cacccctgcg
     2521 gggttccccc acgccttcgg cgcggctccc cctccggcaa aaagtggccc ctccggggct
     2581 tgttgatcga ctgcgcggcc ttcggccttg cccaaggtgg cgctgccccc ttggaacccc
     2641 cgcactcgcc gccgtgaggc tcggggggca ggcgggcggg cttcgccctt cgactgcccc
     2701 cactcgcata ggcttgggtc gttccaggcg cgtcaaggcc aagccgctgc gcggtcgctg
     2761 cgcgagcctt gacccgcctt cctcttggtg tccaaccggc aagcgaagcg cgcaggccgc
     2821 aggccggagg cttttcccca gagaaaatta aaaaaattga tggggcaagg ccgcaggccg
     2881 cgcagttgga gccggtgggt atgtggtcga aggctgggta gccggtgggc aatccctgtg
     2941 gtcaagctcg tgggcaggcg cagcctgtcc atcagcttgt ccagcagggt tgtccacggg
     3001 ccgagcgaag cgagccagcc ggtggccgct cgcggccatc gtccacatat ccacgggctg
     3061 gcaagggagc gcagcgaccg cgcagggcga agcccggaga gcaagcccgt agggggttac
     3121 tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     3181 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctgccg aattcggatc
     3241 cggagtgaga ccgcagctgg cacgacaggt ttgccgactg gaaagcgggc agtgagcgca
     3301 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     3361 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
