LOCUS       sequence-224275-        3135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 high copy number vector for E. coli expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3135)
  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 3135)
  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: pOGG005
FEATURES             Location/Qualifiers
     source          1..3135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    487..508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(523..528,529..546,547..553)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    561..577
                     /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     566..588
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     585..601
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             597..920
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    614..670
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(671..687)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(896..915)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     terminator      1025..1119
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1278..1295)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1388..1976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1468..1487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2147..2938,2939..3007))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2770..2789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3008..3112)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
      361 aggccacgtg tcttgtccag gcgcgccaca ctctgtgcgt ctcatgccga attcggatcc
      421 ggagtgagac cgcagctggc acgacaggtt tgccgactgg aaagcgggca gtgagcgcaa
      481 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
      541 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
      601 ccatgattac gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga
      661 gctcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
      721 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
      781 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
      841 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
      901 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
      961 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacg gtctcacgct
     1021 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
     1081 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggcaatga gacgcacgaa
     1141 gtgattaatt aatggagcac aagactggcc tcatgggcct tccgctcact gcccgctttc
     1201 cagtcgggaa acctgtcgtg ccagctgcat taacatggtc atagctgttt ccttgcgtat
     1261 tgggcgctct ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg ggtaaagcct
     1321 ggggtgccta atgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     2041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     2101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     2161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     2221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     2281 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     2341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     2401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     2461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     2521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     2581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     3001 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     3061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     3121 cccgaaaagt gccac
//
