LOCUS       Exported                5187 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Beta lactamase S68TAG gene.
ACCESSION   .
VERSION     .
KEYWORDS    Beta lactamase S68TAG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5187)
  AUTHORS   Pirman NL, Barber KW, Aerni HR, Ma NJ, Haimovich AD, Rogulina S, 
            Isaacs FJ, Rinehart J
  TITLE     A flexible codon in genomically recoded Escherichia coli permits 
            programmable protein phosphorylation.
  JOURNAL   Nat Commun. 2015 Sep 9;6:8130. doi: 10.1038/ncomms9130.
  PUBMED    26350500
REFERENCE   2  (bases 1 to 5187)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5187
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(1..624)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     532..551
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    669..685
                     /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)."
     protein_bind    692..708
                     /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)."
     promoter        717..790
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    717..735
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    742..760
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             800..811
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             818..1531
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(848..876)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1471..1491
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             1538..1555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1631..1649)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1645..1692
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1789..2244
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1876..1895)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2086..2107
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2270..2374
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2375..3235
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMM*TFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2593..2612)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     primer_bind     3895..3914
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4145..4162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(4411..5070)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4985..5004
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(5071..5173)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 ttaagaccca ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc
       61 gaataagaag gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa
      121 tggcggcata ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg
      181 atcttccaat acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt
      241 ctctagtgaa aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg
      301 tttttctgta ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc
      361 acatctaaaa cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg
      421 gcaaaagtga gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg
      481 cttatttttt acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt
      541 acgggttgtt aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac
      601 tttactttta tctaatctgg acatgtcgaa tatagacgtc agtgcgtcct gctgatgtgc
      661 tcagtatctt gttatccgct cacaatgtca attgttatcc gctcacaatt ctcgagtccc
      721 tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac atcagcagga
      781 cgcactgacc gaattcatta aagaggagaa aggtaccatg agtaaaggag aagaactttt
      841 cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc
      901 tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg
      961 cactactgga aaactacctg ttccatggcc aacacttgtc actactttct cttatggtgt
     1021 tcaatgcttt tcccgttatc cggatcacat gaaacggcat gactttttca agagtgccat
     1081 gcccgaaggt tatgtacagg aacgcactat atctttcaaa gatgacggga actacaagac
     1141 gcgtgctgaa gtcaagtttg aaggtgatac ccttgttaat cgtatcgagt taaaaggtat
     1201 tgattttaaa gaagatggaa acattctcgg acacaaactc gaatacaact ataactcaca
     1261 caatgtatac atcacggcag acaaacaaaa gaatggaatc aaagctaact tcaaaattcg
     1321 ccacaacatt gaagatggat ccgttcaact agcagaccat tatcaacaaa atactccaat
     1381 tggcgatggc cctgtccttt taccagacaa ccattacctg tcgacacaat ctgccctttc
     1441 gaaagatccc aacgaaaagc gtgaccacat ggtccttctt gagtttgtaa ctgctgctgg
     1501 gattacacat ggcatggatg agctctacaa actcgagcac caccaccacc accactgaag
     1561 ggcgagctca attcaaagct tgatccggct gctaacaaag cccgaaagga agctgagttg
     1621 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1681 aggggttttt tgctgaaagg aggaactata tccggatctg gcgtaatagc gaagaggccc
     1741 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     1801 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1861 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1921 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1981 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2041 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2101 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2161 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     2221 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2281 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2341 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2401 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2461 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2521 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgtagac
     2581 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2641 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2701 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2761 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2821 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2881 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     2941 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3001 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3061 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3121 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3181 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3241 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3301 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     3361 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     3421 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     3481 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     3541 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3601 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3661 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     3721 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3781 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3841 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3901 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3961 gtgatgctcg tcaggggctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4021 ccgtattacc gcctttgagt gagctgatac cgctcgggcg gagcctatgg aaacgccagc
     4081 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttccgcagcc gaacgaccga
     4141 gcgcagcgag tcagtgagcg aggaagccaa agggcccaag gcccagtctt tcgactgagc
     4201 ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagac cccacactac
     4261 catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg accaccgcgc
     4321 tactgccgcc aggcatattc atccgcttat tatcacttat tcaggcgtag caccaggcgt
     4381 ttaagggcac caataactgc cttaaaaaaa ttacgccccg ccctgccact catcgcagta
     4441 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacagacgg catgatgaac
     4501 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatggtgaa
     4561 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     4621 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     4681 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     4741 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     4801 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaattccgg
     4861 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     4921 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     4981 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     5041 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     5101 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     5161 tacgtgccga tcaacgtctc actagag
//
