LOCUS       Exported                8580 bp ds-DNA     circular SYN 13-MAY-2021
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    pPaGE Pyl TAG FliC T248TAG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8580)
  TITLE     An inside look at a biofilm: Pseudomonas aeruginosa flagella 
            bio-tracking
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8580)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8580
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(5006..5594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5086..5105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5765..6625)
                     /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     6388..6407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6626..6730)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6798..6816
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6854..6876)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6976..6995
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7716..8067
                     /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)"
     CDS             8081..334
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pRO1600 from Pseudomonas aeruginosa "
                     /label=pRO1600 Rep
                     /translation="MASPPMVYKSNALVEAAYRLSVQEQRIVLACISQVKRSEPVTDEV
                     MYSVTAEDIATMAGVPIESSYNQLKEAALRLKRREVRLTQEPNGKGKRPSVMITGWVQT
                     IIYREGEGRVELRFTKDMLPYLTELTKQFTKYALADVAKMDSTHAIRLYELLMQWDSIG
                     QREIEIDQLRKWFQLEGRYPSIKDFKLRVLDPAVTQINEHSPLQVEWAQRKTGRKVTHL
                     LFSFGPKKPAKAVGKAPAKRKAGKISDAEIAKQARPGETWEAARARLTQMPLDLA"
ORIGIN
        1 agaaattgac cagctgcgaa agtggtttca actggaaggc cggtatccct cgatcaagga
       61 cttcaagttg cgagtgcttg atccagccgt gacgcagatc aacgagcaca gcccgctaca
      121 ggtggagtgg gcgcagcgaa agaccgggcg caaggtcaca catctgttgt tcagttttgg
      181 accgaagaag cccgccaagg cggtgggtaa ggccccagcg aagcgcaagg ccgggaagat
      241 ttcagatgct gagatcgcga aacaggctcg ccctggtgag acatgggaag cggcccgcgc
      301 tcgactaacc cagatgccgc tggatctggc ctagaggccg tggccaccac ggcccggcct
      361 gcctttcgga tcctagaggt aggtcacctt ttcttcttta gcgtttgctg ccattgctct
      421 attgaagcgc gaaaacgctt cgtcggtcag cttcggcaac agaataattt ttatcgcatc
      481 aggggtttac aagcagaata cgtactcgta tagttcgcgc ccacaacgtt gctgagacgt
      541 tgtaacacgg aaacgatgag cgatcatcga ctccagggaa acctgatcat gtagatcgaa
      601 tggactctaa atccgttcag ccgggttaga ttcccggggt ttccgccaat atttcagaca
      661 aagccggtca ttagaccgtt tttgtttcag cagttcagcc actaccaggc attgaaactg
      721 cgacaaggtt tgcaacaccc aagtcgatga gcgatcatcg cacaatgctg aaaagcacga
      781 acacctctga aggtaccact gggcagccag ccatcgacgg cccatagggc aagcttagcg
      841 ttcccgttcg gctgaaccgc ccccggcgac aaacggcgcg ggctatcgac aggagtgggg
      901 gtcatgtatg ctacgaggct ttcgaacagt cacgtctaag tagcgccccg cggatggata
      961 aaaaaccact aaacactctg atctctgcta ctggtctgtg gatgagtcgt accggaacca
     1021 ttcataaaat caaacaccac gaggttagcc gttcgaaaat ctatattgag atggcgtgtg
     1081 gcgatcatct ggttgtgaac aatagccgct cttctcgtac agcacgtgca ctgcgtcacc
     1141 acaaatatcg taaaacctgt aaacgttgcc gtgtgtccga tgaggatctg aacaaattcc
     1201 tgacaaaagc caatgaggac caaacaagcg tgaaagtgaa agtcgttagc gctcctaccc
     1261 gtactaaaaa agcaatgccg aaatccgttg ctcgtgcccc taaaccactg gaaaacactg
     1321 aagcagcaca ggcacagccg tctggaagca aattctctcc ggccattcct gtttctaccc
     1381 aggagtccgt ttctgttcca gcaagtgtga gcaccagcat tagcagtatt agcaccggtg
     1441 ccaccgctag cgccctggtt aaaggcaata ccaatccgat tacaagcatg tctgccccgg
     1501 ttcaagcatc agctccagca ctgacaaaat cccaaaccga tcgtctggag gttctgctga
     1561 atccgaaaga cgaaatcagc ctgaattccg gcaaaccgtt tcgtgaactg gagagcgaac
     1621 tgctgtcacg tcgtaaaaaa gacctgcaac aaatctatgc cgaagaacgt gagaactatc
     1681 tggggaaact ggaacgtgaa atcacccgct ttttcgtgga tcgtggcttt ctggagatca
     1741 aatccccgat tctgattcct ctggagtata tcgagcgtat gggcatcgac aatgataccg
     1801 aactgagcaa acaaattttc cgtgtggata aaaacttctg tctgcgccct atgctggcac
     1861 caaatctgta taactatctg cgcaaactgg accgtgccct gcctgatcct atcaaaatct
     1921 tcgagatcgg cccgtgttat cgtaaagagt ccgacggtaa agaacatctg gaggagttta
     1981 ccatgctgaa cttttgccaa atgggttcag gttgtactcg tgagaacctg gaaagcatca
     2041 tcaccgattt tctgaaccac ctgggcattg acttcaaaat tgtgggcgac agctgtatgg
     2101 tgtatggcga caccctggat gtcatgcacg gcgacctgga actgtctagt gccgttgttg
     2161 gaccaattcc gctggaccgt gagtggggta tcgacaaacc gtggatcgga gcaggattcg
     2221 gtctggaacg cctgctgaaa gtgaaacacg acttcaaaaa catcaaacgt gccgcccgtt
     2281 ctgaatcgta ttataacggg atttctacca acctgtaacc gcggtacgcg attccattgg
     2341 acgcccccgg ccatccgggg gcgctgatca acaggagtga gggaagcttg atccgggttt
     2401 ttctcgaacg aggccggtgg cgcaagccat cggccttttt tcatgcccgg cgtgccctgt
     2461 tgcacgggag ggctaaagaa aatcgccggg gggtcgatgc aatgggtgtc ggaacttcca
     2521 ccctctgccg gaccaaccgg gggcggttca ggaccgatat tggcgagtcc tcttcgaagc
     2581 atgtaaccca ctgaagagga agagaaaaag aaaatgttga ttttttctct aaagctccgc
     2641 cgggaaacgc cgataaacac catgaacgcg aattcttggg gcacctgagc aagcaggccg
     2701 agagatcgca agctcaggta accgaaatag gtcctttgga ggaaatcacc atggccctta
     2761 cagtcaacac gaacattgct tccctgaaca ctcagcgcaa cctgaatgct tcttccaacg
     2821 acctcaacac ctcgttgcag cgtctgacca ccggctaccg catcaacagt gccaaggacg
     2881 atgctgccgg cctgcagatc tccaaccgcc tgtccaacca gatcagcggt ctgaacgttg
     2941 ccacccgcaa cgccaacgac ggcatctccc tggcgcagac cgctgaaggt gccctgcagc
     3001 agtccaccaa tatcctgcag cgtatccgcg acctggccct gcaatccgcc aacggctcca
     3061 acagcgacgc cgaccgtgcc gccctgcaga aagaagtcgc tgcgcaacag gccgaactga
     3121 cccgtatctc cgataccacc accttcggtg gccgcaagct gctcgacggc tccttcggca
     3181 ccaccagctt ccaggtcggt tccaacgcct acgagaccat tgacatcagc ctgcagaatg
     3241 cctctgccag cgccatcggt tcttaccagg tcggcagcaa cggcgcgggt accgtcgcca
     3301 gcgtagcggg caccgcgacc gcttcgggca tcgcctcggg caccgtcaac ctggtcggtg
     3361 gcggtcaggt gaagaacatc gccatcgccg ccggcgatag cgccaaggcc atcgccgaga
     3421 agatggacgg tgcgatcccg aacctgtcgg ctcgtgcccg taccgtgttc accgctgatg
     3481 tcagcggcgt gtagggtggt tcgctgaact tcgacgtaac cgttggcagc aacaccgtga
     3541 gcctggcagg cgtgacctcc actcaggatc tggccgacca actgaactcc aactcgtcga
     3601 agctgggcat cactgccagc atcaacgaca agggtgtact gaccatcacc tccgctaccg
     3661 gcgagaacgt caagttcggt gcgcagaccg gtaccgctac tgccggtcag gtcgcagtga
     3721 aggtccaggg ttccgacggc aagttcgaag cggccgccaa gaacgtggta gctgccggta
     3781 ctgccgctac caccaccatc gtgaccggct acgtgcaact gaactcgccg accgcctact
     3841 cggtcagcgg taccggcacc caggcttcgc aggtcttcgg caacgccagc gccgcgcaga
     3901 agagcagcgt tgccagcgtc gacatctcca ctgccgacgg cgcccagaac gccatcgcgg
     3961 tagtcgataa cgccctggct gcgatcgacg cccagcgtgc tgacctcggt gctgttcaga
     4021 accgcttcaa gaacactatc gacaacctga ccaacatctc ggaaaacgct accaacgctc
     4081 gtagccgcat caaggacacc gacttcgctg ccgaaaccgc ggcgctgtcg aagaaccagg
     4141 tgctgcaaca ggccggtacc gcgatcctgg cccaggccaa ccagctgccg caggcggtcc
     4201 tgagcctgct gcgctaagcc cgggaacggt cactcacgcg gtactgggag gaaggggtga
     4261 cccttcctcc cttttcccca tggagggcac agttaagccg ctaaaggcat tatccgccaa
     4321 gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata cagtcaaatt
     4381 gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga atgcacacgg
     4441 tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag taacagagga
     4501 acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat ctactggaga
     4561 gtatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta tcggctttat
     4621 tgctcaaaga gacatgggtg gaagagatga aggttacgat tggctgatta tgacacccgg
     4681 tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg tggatgatgt
     4741 ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa agggaaggga
     4801 tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt tgagaagatg
     4861 cggccagcaa aactactagt gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4921 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4981 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5041 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5101 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5161 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5221 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5281 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5341 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5401 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5461 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5521 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5581 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5641 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5701 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5761 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5821 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5881 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5941 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6001 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6061 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6121 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6181 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6241 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6301 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6361 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6421 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6481 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6541 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6601 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6661 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6721 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6781 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6841 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6901 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6961 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7021 aaataccgca cagatgcgta aggagaaaat accgcatcag gcggcaatgg caacaacgtt
     7081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7201 tattgctgat aaatctggag ccggtgagcg tggatctcgc ggtatcattg cagcactggg
     7261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7501 ttcgttccac tgagcgtcag accccaatta cacgccactg gctgtgcttg ctggggtgac
     7561 ggtggcaacg gtggcggcct tgctgggcta tcgcgttgga aagaaacgag ggaaagggga
     7621 ctgataaacc ggtcttagcc cctccccttg gtgtccaacc gctctgtagg cctctcaggc
     7681 gccgctggtg ccgctggttg gacgccaagg gtgaatccgc ctcgataccc tgattactcg
     7741 cttcctgcgc cctctcaggc ggcgataggg gactggtaaa acggggattg cccagacgcc
     7801 tcccccgccc cttcaggggc acaaatgcgg ccccaacggg gccacgtagt ggtgcgtttt
     7861 ttgcgtttcc acccttttct tccttttccc ttttaaacct tttaggacgt ctacaggcca
     7921 cgtaatccgt ggcctgtaga gtttaaaaag ggacggattt gttgccatta agggacggat
     7981 ttgttgttaa gaagggacgg atttgttgtt gtaaagggac ggatttgttg tattgtggga
     8041 cgcagataca gtgtcccctt atacacaagg aatgtcgaac gtggcctcac ccccaatggt
     8101 ttacaaaagc aatgccctgg tcgaggccgc gtatcgcctc agtgttcagg aacagcggat
     8161 cgttctggcc tgtattagcc aggtgaagag gagcgagcct gtcaccgatg aagtgatgta
     8221 ttcagtgacg gcggaggaca tagcgacgat ggcgggtgtc cctatcgaat cttcctacaa
     8281 ccagctcaaa gaagcggccc tgcgcctgaa acggcgggaa gtccggttaa cccaagagcc
     8341 caatggcaag gggaaaagac cgagtgtgat gattaccggc tgggtgcaaa caatcatcta
     8401 ccgggagggt gagggccgtg tagaactcag gttcaccaaa gacatgctgc cgtacctgac
     8461 ggaactcacc aaacagttca ccaaatacgc cttggctgac gtggccaaga tggacagcac
     8521 ccacgcgatc aggctttacg agctgctcat gcaatgggac agcatcggcc agcgcgaaat
//
