LOCUS       sequence-310028-        8660 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for MmPyl tRNA synthetase/tRNA pair and for flagellin 
            protein (FliC) with TAG mutation at site 248, for unnatural amino 
            acid incorporation into the flagellin protein in Pseudomonas 
            aeruginosa..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8660)
  AUTHORS   Ozer E, Yaniv K, Chetrit E, Boyarski A, Meijler MM, Berkovich R, 
            Kushmaro A, Alfonta L
  TITLE     An inside look at a biofilm: Pseudomonas aeruginosa flagella 
            biotracking.
  JOURNAL   Sci Adv. 2021 Jun 11;7(24). pii: 7/24/eabg8581. doi: 
            10.1126/sciadv.abg8581. Print 2021 Jun.
  PUBMED    34117070
REFERENCE   2  (bases 1 to 8660)
  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: "Sci Adv."; 
            date: "2021-06-11"; volume: "7(24). pii: 7/24/eabg8581. doi"; pages:
            " 10.1126/sciadv.abg8581. Print 2021 Jun"
COMMENT     Sequence Label: pPaGE Pyl TAG FliC T248TAG
FEATURES             Location/Qualifiers
     source          1..8660
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(5086..5674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5166..5185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5845..6636,6637..6705))
                     /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     6468..6487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6706..6810)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6878..6896
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6934..6956)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7056..7075
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7796..8147
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from 
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
ORIGIN
        1 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga tacactagta
       61 gttttgctgg ccgcatcttc agaaattgac cagctgcgaa agtggtttca actggaaggc
      121 cggtatccct cgatcaagga cttcaagttg cgagtgcttg atccagccgt gacgcagatc
      181 aacgagcaca gcccgctaca ggtggagtgg gcgcagcgaa agaccgggcg caaggtcaca
      241 catctgttgt tcagttttgg accgaagaag cccgccaagg cggtgggtaa ggccccagcg
      301 aagcgcaagg ccgggaagat ttcagatgct gagatcgcga aacaggctcg ccctggtgag
      361 acatgggaag cggcccgcgc tcgactaacc cagatgccgc tggatctggc ctagaggccg
      421 tggccaccac ggcccggcct gcctttcgga tcctagaggt aggtcacctt ttcttcttta
      481 gcgtttgctg ccattgctct attgaagcgc gaaaacgctt cgtcggtcag cttcggcaac
      541 agaataattt ttatcgcatc aggggtttac aagcagaata cgtactcgta tagttcgcgc
      601 ccacaacgtt gctgagacgt tgtaacacgg aaacgatgag cgatcatcga ctccagggaa
      661 acctgatcat gtagatcgaa tggactctaa atccgttcag ccgggttaga ttcccggggt
      721 ttccgccaat atttcagaca aagccggtca ttagaccgtt tttgtttcag cagttcagcc
      781 actaccaggc attgaaactg cgacaaggtt tgcaacaccc aagtcgatga gcgatcatcg
      841 cacaatgctg aaaagcacga acacctctga aggtaccact gggcagccag ccatcgacgg
      901 cccatagggc aagcttagcg ttcccgttcg gctgaaccgc ccccggcgac aaacggcgcg
      961 ggctatcgac aggagtgggg gtcatgtatg ctacgaggct ttcgaacagt cacgtctaag
     1021 tagcgccccg cggatggata aaaaaccact aaacactctg atctctgcta ctggtctgtg
     1081 gatgagtcgt accggaacca ttcataaaat caaacaccac gaggttagcc gttcgaaaat
     1141 ctatattgag atggcgtgtg gcgatcatct ggttgtgaac aatagccgct cttctcgtac
     1201 agcacgtgca ctgcgtcacc acaaatatcg taaaacctgt aaacgttgcc gtgtgtccga
     1261 tgaggatctg aacaaattcc tgacaaaagc caatgaggac caaacaagcg tgaaagtgaa
     1321 agtcgttagc gctcctaccc gtactaaaaa agcaatgccg aaatccgttg ctcgtgcccc
     1381 taaaccactg gaaaacactg aagcagcaca ggcacagccg tctggaagca aattctctcc
     1441 ggccattcct gtttctaccc aggagtccgt ttctgttcca gcaagtgtga gcaccagcat
     1501 tagcagtatt agcaccggtg ccaccgctag cgccctggtt aaaggcaata ccaatccgat
     1561 tacaagcatg tctgccccgg ttcaagcatc agctccagca ctgacaaaat cccaaaccga
     1621 tcgtctggag gttctgctga atccgaaaga cgaaatcagc ctgaattccg gcaaaccgtt
     1681 tcgtgaactg gagagcgaac tgctgtcacg tcgtaaaaaa gacctgcaac aaatctatgc
     1741 cgaagaacgt gagaactatc tggggaaact ggaacgtgaa atcacccgct ttttcgtgga
     1801 tcgtggcttt ctggagatca aatccccgat tctgattcct ctggagtata tcgagcgtat
     1861 gggcatcgac aatgataccg aactgagcaa acaaattttc cgtgtggata aaaacttctg
     1921 tctgcgccct atgctggcac caaatctgta taactatctg cgcaaactgg accgtgccct
     1981 gcctgatcct atcaaaatct tcgagatcgg cccgtgttat cgtaaagagt ccgacggtaa
     2041 agaacatctg gaggagttta ccatgctgaa cttttgccaa atgggttcag gttgtactcg
     2101 tgagaacctg gaaagcatca tcaccgattt tctgaaccac ctgggcattg acttcaaaat
     2161 tgtgggcgac agctgtatgg tgtatggcga caccctggat gtcatgcacg gcgacctgga
     2221 actgtctagt gccgttgttg gaccaattcc gctggaccgt gagtggggta tcgacaaacc
     2281 gtggatcgga gcaggattcg gtctggaacg cctgctgaaa gtgaaacacg acttcaaaaa
     2341 catcaaacgt gccgcccgtt ctgaatcgta ttataacggg atttctacca acctgtaacc
     2401 gcggtacgcg attccattgg acgcccccgg ccatccgggg gcgctgatca acaggagtga
     2461 gggaagcttg atccgggttt ttctcgaacg aggccggtgg cgcaagccat cggccttttt
     2521 tcatgcccgg cgtgccctgt tgcacgggag ggctaaagaa aatcgccggg gggtcgatgc
     2581 aatgggtgtc ggaacttcca ccctctgccg gaccaaccgg gggcggttca ggaccgatat
     2641 tggcgagtcc tcttcgaagc atgtaaccca ctgaagagga agagaaaaag aaaatgttga
     2701 ttttttctct aaagctccgc cgggaaacgc cgataaacac catgaacgcg aattcttggg
     2761 gcacctgagc aagcaggccg agagatcgca agctcaggta accgaaatag gtcctttgga
     2821 ggaaatcacc atggccctta cagtcaacac gaacattgct tccctgaaca ctcagcgcaa
     2881 cctgaatgct tcttccaacg acctcaacac ctcgttgcag cgtctgacca ccggctaccg
     2941 catcaacagt gccaaggacg atgctgccgg cctgcagatc tccaaccgcc tgtccaacca
     3001 gatcagcggt ctgaacgttg ccacccgcaa cgccaacgac ggcatctccc tggcgcagac
     3061 cgctgaaggt gccctgcagc agtccaccaa tatcctgcag cgtatccgcg acctggccct
     3121 gcaatccgcc aacggctcca acagcgacgc cgaccgtgcc gccctgcaga aagaagtcgc
     3181 tgcgcaacag gccgaactga cccgtatctc cgataccacc accttcggtg gccgcaagct
     3241 gctcgacggc tccttcggca ccaccagctt ccaggtcggt tccaacgcct acgagaccat
     3301 tgacatcagc ctgcagaatg cctctgccag cgccatcggt tcttaccagg tcggcagcaa
     3361 cggcgcgggt accgtcgcca gcgtagcggg caccgcgacc gcttcgggca tcgcctcggg
     3421 caccgtcaac ctggtcggtg gcggtcaggt gaagaacatc gccatcgccg ccggcgatag
     3481 cgccaaggcc atcgccgaga agatggacgg tgcgatcccg aacctgtcgg ctcgtgcccg
     3541 taccgtgttc accgctgatg tcagcggcgt gtagggtggt tcgctgaact tcgacgtaac
     3601 cgttggcagc aacaccgtga gcctggcagg cgtgacctcc actcaggatc tggccgacca
     3661 actgaactcc aactcgtcga agctgggcat cactgccagc atcaacgaca agggtgtact
     3721 gaccatcacc tccgctaccg gcgagaacgt caagttcggt gcgcagaccg gtaccgctac
     3781 tgccggtcag gtcgcagtga aggtccaggg ttccgacggc aagttcgaag cggccgccaa
     3841 gaacgtggta gctgccggta ctgccgctac caccaccatc gtgaccggct acgtgcaact
     3901 gaactcgccg accgcctact cggtcagcgg taccggcacc caggcttcgc aggtcttcgg
     3961 caacgccagc gccgcgcaga agagcagcgt tgccagcgtc gacatctcca ctgccgacgg
     4021 cgcccagaac gccatcgcgg tagtcgataa cgccctggct gcgatcgacg cccagcgtgc
     4081 tgacctcggt gctgttcaga accgcttcaa gaacactatc gacaacctga ccaacatctc
     4141 ggaaaacgct accaacgctc gtagccgcat caaggacacc gacttcgctg ccgaaaccgc
     4201 ggcgctgtcg aagaaccagg tgctgcaaca ggccggtacc gcgatcctgg cccaggccaa
     4261 ccagctgccg caggcggtcc tgagcctgct gcgctaagcc cgggaacggt cactcacgcg
     4321 gtactgggag gaaggggtga cccttcctcc cttttcccca tggagggcac agttaagccg
     4381 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac
     4441 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca
     4501 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg
     4561 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag
     4621 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa
     4681 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat
     4741 tggttgatta tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag
     4801 tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga
     4861 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg
     4921 gaagcatatt tgagaagatg cggccagcaa aactactagt gtatcagctc actcaaaggc
     4981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5521 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     6001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     6061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     6121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     6181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6841 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6901 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6961 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     7021 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     7081 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     7141 gcggcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     7201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     7261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggatctcgc
     7321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     7381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     7441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     7501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     7561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccaatta cacgccactg
     7621 gctgtgcttg ctggggtgac ggtggcaacg gtggcggcct tgctgggcta tcgcgttgga
     7681 aagaaacgag ggaaagggga ctgataaacc ggtcttagcc cctccccttg gtgtccaacc
     7741 gctctgtagg cctctcaggc gccgctggtg ccgctggttg gacgccaagg gtgaatccgc
     7801 ctcgataccc tgattactcg cttcctgcgc cctctcaggc ggcgataggg gactggtaaa
     7861 acggggattg cccagacgcc tcccccgccc cttcaggggc acaaatgcgg ccccaacggg
     7921 gccacgtagt ggtgcgtttt ttgcgtttcc acccttttct tccttttccc ttttaaacct
     7981 tttaggacgt ctacaggcca cgtaatccgt ggcctgtaga gtttaaaaag ggacggattt
     8041 gttgccatta agggacggat ttgttgttaa gaagggacgg atttgttgtt gtaaagggac
     8101 ggatttgttg tattgtggga cgcagataca gtgtcccctt atacacaagg aatgtcgaac
     8161 gtggcctcac ccccaatggt ttacaaaagc aatgccctgg tcgaggccgc gtatcgcctc
     8221 agtgttcagg aacagcggat cgttctggcc tgtattagcc aggtgaagag gagcgagcct
     8281 gtcaccgatg aagtgatgta ttcagtgacg gcggaggaca tagcgacgat ggcgggtgtc
     8341 cctatcgaat cttcctacaa ccagctcaaa gaagcggccc tgcgcctgaa acggcgggaa
     8401 gtccggttaa cccaagagcc caatggcaag gggaaaagac cgagtgtgat gattaccggc
     8461 tgggtgcaaa caatcatcta ccgggagggt gagggccgtg tagaactcag gttcaccaaa
     8521 gacatgctgc cgtacctgac ggaactcacc aaacagttca ccaaatacgc cttggctgac
     8581 gtggccaaga tggacagcac ccacgcgatc aggctttacg agctgctcat gcaatgggac
     8641 agcatcggcc agcgcgaaat
//
