LOCUS       sequence-343491-        5838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression protein designed PodA10 from a TEV cleavable IPTG 
            inducible plasmid with ampicillin resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5838)
  AUTHORS   VanDrisse CM, Lipsh-Sokolik R, Khersonsky O, Fleishman SJ, Newman DK
  TITLE     Computationally designed pyocyanin demethylase acts synergistically 
            with tobramycin to kill recalcitrant Pseudomonas aeruginosa 
            biofilms.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Mar 23;118(12). pii: 2022012118. doi:
            10.1073/pnas.2022012118.
  PUBMED    33723058
REFERENCE   2  (bases 1 to 5838)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-03-23"; volume: "118(12). pii: 
            2022012118. doi"; pages: " 10.1073/pnas.2022012118"
COMMENT     Sequence Label: pPodA11
FEATURES             Location/Qualifiers
     source          1..5838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     184..201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    216..355
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(281..300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     441..463
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(457..648)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1457..2539)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2501..2520
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2540..2617)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2823..2842
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2926..2945
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2926..2944
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2945..2969
                     /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)."
     RBS             2984..3006
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3028..3045
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3067..3087
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3484..3502)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3498..3545
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3582..4037
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3669..3688)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3879..3900
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4063..4167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4168..4236,4237..5028)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4386..4405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     5769..5788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
       61 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
      121 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
      181 cgcagcgagt cagtgagcga ggaagcggaa tagagcctga tgcggtattt tctccttacg
      241 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
      301 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
      361 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
      421 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
      481 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
      541 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
      601 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
      661 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
      721 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
      781 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
      841 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
      901 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
      961 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     1021 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     1081 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     1141 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     1201 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     1261 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     1321 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     1381 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     1441 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1501 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt
     1561 ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga
     1621 gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg
     1681 ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat
     1741 ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat
     1801 cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt
     1861 gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc
     1921 gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc
     1981 ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg
     2041 taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa
     2101 ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg
     2161 gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac
     2221 aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg
     2281 cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg
     2341 caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt
     2401 aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg
     2461 cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt
     2521 ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg
     2581 ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta
     2641 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     2701 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     2761 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     2821 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     2881 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
     2941 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
     3001 aggagatata catatatgtc gtactaccat caccatcacc atcacgatta cgatatccca
     3061 actagtgaaa acctgtattt tcagggcatg gttacaatga cgcttgacgt aaagaatgac
     3121 caagtcaaca agcatgattt tggcaagcct ggaatggacg taggtgacat ggacattttc
     3181 tccgatactc tgagcgtaga cgggaagcaa gttgggtatg atggaggagt ctgctttttt
     3241 acgaatgtaa cccctgacaa ccccgtgacc tactgcgagc ttactattca cttagacgca
     3301 ggtgaaattt tcgctcgtag tttgacccca catacattag cgccttttac tatgacaatt
     3361 acagggggga caggcgaata tgcgaactcg aagggtgaac ttacagtgag tggcgtggca
     3421 acccctgacg aaaagtacga gttaaagtta acataagaat tctcgagctc ccgggatccg
     3481 cggccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
     3541 ttttgctgaa aggaggaact atatccggat tggcgaatgg gacgcgccct gtagcggcgc
     3601 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3661 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3721 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3781 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3841 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3901 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     3961 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4021 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4081 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4141 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4201 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4261 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4321 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4381 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4441 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4501 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4561 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4621 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4681 ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta
     4741 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4801 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4861 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4921 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4981 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5041 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5101 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5161 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5221 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5281 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5341 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5401 acatacctcg ctctgctaat cctgttacca gtggatcgac ggagcgatgc cgagccgtgc
     5461 tgaaacggta aaaggttacg tgcaggccat gcaccagagc tggttacagg atgtgctgct
     5521 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5581 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5641 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5701 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5761 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5821 gagcgtcgat ttttgtga
//
