LOCUS       sequence-449823-        5825 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Control biosensor for extracellular L-lactate.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5825)
  TITLE     
REFERENCE   2  (bases 1 to 5825)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pBAD-R-eLACCO2.1
FEATURES             Location/Qualifiers
     source          1..5825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(339..353,354..362))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(366..398)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(402..419)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(465..749)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(523..542)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             776..1654
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(1810..1832)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1918..2058
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1973..1992
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2073..2090)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2244..2832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2324..2343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3003..3794,3795..3863))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3626..3645
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3864..3955)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(3974..4001)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4093..4179)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     4332..4349
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     primer_bind     complement(4861..4880)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 aaccatgttt cccactggtc aggtcggtcc aacccgaggg cgtagctgga gaggaaagct
       61 gtaaccggca tcctctctgc ccaatagatc gtgaaaggat tcattccgtc caaaactcca
      121 gtcttgaccg cgtcgaacat atcaaacgtg cctacgaccg cacccgctgg aagaggctgg
      181 acctcaagct gtccatccgt cagttccttt acccgctccg taaacttttg gaacagactg
      241 tagcctactg ttccagcatc ccacgcggtt tgaatcctcc acttatagcg tcgagcctgg
      301 gccactgcca tgggagagaa cattctcgag ctcggatcct tatcgtcatc gtcgtacaga
      361 tcccgaccca tttgctgtcc accagtcatg ctagccatac catgatgatg atgatgatga
      421 gaacccccca tggttaattc ctcctgttag cccaaaaaac gggtatggag aaacagtaga
      481 gagttgcgat aaaaagcgtc aggtaggatc cgctaatctt atggataaaa atgctatggc
      541 atagcaaagt gtgacgccgt gcaaataatc aatgtggact tttctgccgt gattatagac
      601 acttttgtta cgcgtttttg tcatggcttt ggtcccgctt tgttacagaa tgcttttaat
      661 aagcggggtt accggtttgg ttagcgagaa gagccagtaa aagacgcagt gacggcaatg
      721 tctgatgcaa tatggacaat tggtttcttc tctgaatggc gggagtatga aaagtatggc
      781 tgaagcgcaa aatgatcccc tgctgccggg atactcgttt aatgcccatc tggtggcggg
      841 tttaacgccg attgaggcca acggttatct cgattttttt atcgaccgac cgctgggaat
      901 gaaaggttat attctcaatc tcaccattcg cggtcagggg gtggtgaaaa atcagggacg
      961 agaatttgtt tgccgaccgg gtgatatttt gctgttcccg ccaggagaga ttcatcacta
     1021 cggtcgtcat ccggaggctc gcgaatggta tcaccagtgg gtttactttc gtccgcgcgc
     1081 ctactggcat gaatggctta actggccgtc aatatttgcc aatacggggt tctttcgccc
     1141 ggatgaagcg caccagccgc atttcagcga cctgtttggg caaatcatta acgccgggca
     1201 aggggaaggg cgctattcgg agctgctggc gataaatctg cttgagcaat tgttactgcg
     1261 gcgcatggaa gcgattaacg agtcgctcca tccaccgatg gataatcggg tacgcgaggc
     1321 ttgtcagtac atcagcgatc acctggcaga cagcaatttt gatatcgcca gcgtcgcaca
     1381 gcatgtttgc ttgtcgccgt cgcgtctgtc acatcttttc cgccagcagt tagggattag
     1441 cgtcttaagc tggcgcgagg accaacgtat cagccaggcg aagctgcttt tgagcaccac
     1501 ccggatgcct atcgccaccg tcggtcgcaa tgttggtttt gacgatcaac tctatttctc
     1561 gcgggtattt aaaaaatgca ccggggccag cccgagcgag ttccgtgccg gttgtgaaga
     1621 aaaagtgaat gatgtagccg tcaagttgtc ataattggta acgaatcaga caattgacgg
     1681 cttgacggag tagcataggg tttgcagaat ccctgcttcg tccatttgac aggcacatta
     1741 tgcatgccgc ttcgccttcg cgcgcgaatt gatctgctgc ctcgcgcgtt tcggtgatga
     1801 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     1861 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc
     1921 agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca
     1981 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2041 agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2101 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2161 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2221 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2281 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2341 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2401 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2461 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2521 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2581 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2641 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     2701 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2761 caaaccaccg ctggtagcgg tggttgtttt gtttgcaagc agcagattac gcgcagaaaa
     2821 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2881 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2941 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     3001 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     3061 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     3121 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     3181 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     3241 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     3301 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     3361 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     3421 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     3481 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3541 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3601 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     3661 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3721 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3781 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3841 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     3901 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt
     3961 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc
     4021 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc
     4081 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     4141 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     4201 tcgcatgggg agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca
     4261 tggggtcagg tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc
     4321 ttctgcgttc tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc
     4381 caagctttca caggctcaag cccttcaatt cctcgtccgt aacgtgacct atacctctca
     4441 tatactccag ttgggacgcg aaagcctctc gtgagtattt atccatctta gcccacttaa
     4501 accaaatggg gatagcgagt cgcctgaatt ttctaacatc ctcgttgccc agccggatga
     4561 cctcaacacc tgcctgccga tatttcggcc acgcctccaa attagccttc tgaatgcctg
     4621 cgtagtgaat ccaagagtac tcgtgaaccg ccgcgatgaa tctttcttgc agaggctttg
     4681 gaaggctgcg ccaccgatta aggtttatgg tgaagtccat aaggtctaca ggttgatgaa
     4741 tggcaggcgt ctctggaggc cccattatga tgtactttgc ctcctgatgg aaccccaagt
     4801 tatagtttac tgcaggccca acaaaatcgg ccgcactacg cccccgctcc cagcccatgg
     4861 tcttcttctg cattacgggg ccgtcggggg ggaagttggt gccgcgcagc ttcaccttgt
     4921 agatgaatac gccgtcctgc agggaggagt cctggtcaac gtgaataatg ccgccgtcct
     4981 cgaagtacat cacgcgctcc cacctgaagc cctcggggaa ggacagcttg aagtagtcgg
     5041 ggatgtcggc tgggtgctta atgtaggcct tggagccgta catgaactga ggggatagga
     5101 tgtcccaggc gaagggcagg gggccaccct tggtcacctt cagcttagcg gtctgaaagg
     5161 cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa ctcgtggccg ttcacggagc
     5221 cctccatgtg caccttgaag cgcatgaact ccttgacgat ggccatgtca tcctcctcgc
     5281 ccttgctcac cagactcccg cctgtacctc ccttgtccag ctcgaccacg ccgccggtgg
     5341 agtggcggcc ctcggcgcgt tcgcactgtt ccacgatagt gtagtcctcg ttgtgggaca
     5401 cgatgtccaa ctggatgtcg acgacgtagg cgccgggcag ctgcacgggc ttcttggcct
     5461 tgtaggtggt cttgaccacg gcggcgtagt ggccgccgtc cttcagcctc agccccttct
     5521 tgatctcgct cttcagggcg ccgtcctcgg ggtacatccg ctcggaaacc gcagtcactg
     5581 cccctctttc cagagcgggg tagacctccc ccccaggcaa gaggacggtg gaagctcctg
     5641 cggccgcaaa gacttccgca atcatgtcgc ctggcactcg gagcttgaca cctttgaagt
     5701 cctcaaatct ccggatcggc ttctttgaat gaattgtatt gaggtcgtgc tgaactggtc
     5761 ccacgtagaa gaggccttgc tcggcaaagg ctcgtcgggc gttgtcgagg ccacccaggg
     5821 agtaa
//
