LOCUS       sequence-361616-        6893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  the patatin-like phosphodiesterase CapV variant CapVQ329R cloned in 
            the L-arabinose inducible vector pBAD28 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6893)
  AUTHORS   Li F, Cao L, Bahre H, Kim SK, Schroeder K, Jonas K, Koonce K, 
            Mekonnen SA, Mohanty S, Bai F, Brauner A, Lee VT, Rohde M, Romling U
  TITLE     Patatin-like phospholipase CapV in Escherichia coli - morphological 
            and physiological effects of one amino acid substitution.
  JOURNAL   NPJ Biofilms Microbiomes. 2022 May 11;8(1):39. doi: 
            10.1038/s41522-022-00294-z.
  PUBMED    35546554
REFERENCE   2  (bases 1 to 6893)
  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: "NPJ 
            Biofilms Microbiomes."; date: "2022-05-11"; volume: "8(1):39. doi"; 
            pages: " 10.1038/s41522-022-00294-z"
COMMENT     Sequence Label: pCapVQ329R
FEATURES             Location/Qualifiers
     source          1..6893
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /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     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     regulatory      2142..2151
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2482..2499)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2652..2738
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2830..2857
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2876..2967
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2968..3036,3037..3828)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3186..3205)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3870..4325
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3957..3976)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4167..4188
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4881..5540)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5455..5474
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5541..5643)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(6035..6052)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6169..6714)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(6249..6268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6790..6809
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcgaatt cgagctcgct
     1321 atattctctg gttatggggt tttcaatgtc tgaattgcat gtagtcaatg accatggagt
     1381 acggctactc tgcctaaatg gtggtggggc cagagggatg tttacaatca gtgtgcttgc
     1441 tgagattgag aggatacttg cctctaggca ccctgaccaa gaaatcagaa ttggcgatta
     1501 ctttgacctc atcgcaggga catcgatagg tgggatcttg gctctcggcc ttgctaaagg
     1561 caagagtgct cgcgagttgg aatgtgtttt cttagataaa gcgaatgata tttttccacc
     1621 aagatggtgg ctattcaatc tccttaaatc acttgtttct cccatttata gtagtaaacc
     1681 acttcgacaa acaattgaaa cgatgatcgg tggtgagact acctttaatg acctggcccg
     1741 gagagtaatg gtcccggcag tcaatctttc aacaggaaaa cctcagttct tcaagacacc
     1801 acataaccca gattttaccc gcgacggtgc acttaaattg attgatgccg cactcgcaac
     1861 ctccgctgca ccgacgttct ttgcaccaca ccattgtgaa gaccttggtt cttattttgc
     1921 tgatggaggt ctggtcgcca acaaccccag ttatatcgga tttttggaag ttttcaggga
     1981 catgaagtct gattttcctg atgttgctca taaagacata cacatactta atatcggtac
     2041 taccggggaa gagtattcac ttagcccaag gctgctgagc aaaaaatggt ggaatggcta
     2101 ctgccgactt tggggcgtag gtaaaaaact ggtccttaca accatggcgg ctaaccaaca
     2161 tctccataaa aatatgctcc tgcgcgagct tacgctccat ggtgcctccg ataactacac
     2221 cagtctggat gaagtgatcc ctaatgaagc agccagcgac atcactctgg ataatgccac
     2281 gaagagttcc attgaaaacc tgcgcgcaag gggaagacga ctcgctactg tgcagttcgc
     2341 tcagaaccag aagcttaaga gctttttcac tacacctgca aaacccttta agcgcgcaat
     2401 cgcgcaggag aaactctgat ctagagtcga cctgcaggca tgcaagcttg gctgttttgg
     2461 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     2521 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     2581 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     2641 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2701 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     2761 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     2821 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     2881 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2941 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3001 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     3061 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     3121 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     3181 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     3241 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     3301 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     3361 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     3421 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3481 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     3541 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3601 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3661 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3721 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3781 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3841 gtttactcat atatacttta gattgattta cgcgccctgt agcggcgcat taagcgcggc
     3901 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3961 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4021 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4081 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4141 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4201 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4261 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     4321 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4381 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4441 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4501 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     4561 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4621 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtc aggcatttga
     4681 gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc aatagacata
     4741 agcggctatt taacgaccct gccctgaacc gacgaccggg tcgaatttgc tttcgaattt
     4801 ctgccattca tccgcttatt atcacttatt caggcgtagc aaccaggcgt ttaagggcac
     4861 caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta ctgttgtaat
     4921 tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc
     4981 agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa aacgggggcg
     5041 aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac ccagggattg
     5101 gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag gttttcaccg
     5161 taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc gtggtattca
     5221 ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca
     5281 ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg atgagcattc
     5341 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
     5401 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
     5461 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
     5521 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
     5581 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
     5641 tcaacgtctc attttcgcca aaagttggcc cagggcttcc cggtatcaac agggacacca
     5701 ggatttattt attctgcgaa gtgatcttcc gtcacaggta tttattcggc gcaaagtgcg
     5761 tcgggtgatg ctgccaactt actgatttag tgtatgatgg tgtttttgag gtgctccagt
     5821 ggcttctgtt tctatcagct gtccctcctg ttcagctact gacggggtgg tgcgtaacgg
     5881 caaaagcacc gccggacatc agcgctagcg gagtgtatac tggcttacta tgttggcact
     5941 gatgagggtg tcagtgaagt gcttcatgtg gcaggagaaa aaaggctgca ccggtgcgtc
     6001 agcagaatat gtgatacagg atatattccg cttcctcgct cactgactcg ctacgctcgg
     6061 tcgttcgact gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg
     6121 ccaggaagat acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct
     6181 ccgcccccct gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac
     6241 aggactataa agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc
     6301 tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc
     6361 tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg
     6421 ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac
     6481 atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag
     6541 tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc
     6601 cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag
     6661 gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat
     6721 catcttatta atcagataaa atatttctag gctcatgagc ccgaagtggc gagcccgatc
     6781 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     6841 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atc
//
