LOCUS       Exported                6201 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses Cph8, Ho1, and PcyA constitutively.
ACCESSION   .
VERSION     .
KEYWORDS    pSR33.4r
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6201)
  AUTHORS   Schmidl SR, Sheth RU, Wu A, Tabor JJ
  TITLE     Refactoring and optimization of light-switchable escherichia coli 
            two-component systems.
  JOURNAL   ACS Synth Biol. 2014 Nov 21;3(11):820-31. doi: 10.1021/sb500273n. 
            Epub 2014 Oct 11.
  PUBMED    25250630
REFERENCE   2  (bases 1 to 6201)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6201
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        2..35
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             52..63
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     RBS             795..806
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1574..1645
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1661..1688
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      4051..4137
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             4305..5096
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     terminator      5108..5202
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5316..5861
                     /direction=RIGHT
                     /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     5762..5781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5978..5995
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctgacagcta gctcagtcct aggtataatg ctagctcaca cagaattcat taaagaggag
       61 aaaggtacca tgagtgtcaa cttagcttcc cagttgcggg aagggacgaa aaaatcccac
      121 tccatggcgg agaacgtcgg ctttgtcaaa tgcttcctca agggcgttgt cgagaaaaat
      181 tcctaccgta agctggttgg caatctctac tttgtctaca gtgccatgga agaggaaatg
      241 gcaaaattta aggaccatcc catcctcagc cacatttact tccccgaact caaccgcaaa
      301 caaagcctag agcaagacct gcaattctat tacggctcca actggcggca agaagtgaaa
      361 atttctgccg ctggccaagc ctatgtggac cgagtccggc aagtggccgc tacggcccct
      421 gaattgttgg tggcccattc ctacacccgt tacctggggg atctttccgg cggtcaaatt
      481 ctcaagaaaa ttgcccaaaa tgccatgaat ctccacgatg gtggcacagc tttctatgaa
      541 tttgccgaca ttgatgacga aaaggctttt aaaaatacct accgtcaagc tatgaatgat
      601 ctgcccattg accaagccac cgccgaacgg attgtggatg aagccaatga cgcctttgcc
      661 atgaacatga aaatgttcaa cgaacttgaa ggcaacctga tcaaggcgat cggcattatg
      721 gtgttcaaca gcctcacccg tcgccgcagt caaggcagca ccgaagttgg cctcgccacc
      781 tccgaaggct agttaaagag gagaaaggat ccatggccgt cactgattta agtttgacca
      841 attcttccct gatgcccacg ttgaacccga tgattcaaca gttggccctg gcgatcgccg
      901 ctagttggca aagtttaccc ctcaagccct atcaattgcc ggaggatttg ggctacgtag
      961 aaggccgcct ggaaggggaa aagttagtga ttgaaaatcg gtgctaccaa acgccccagt
     1021 ttcgcaaaat gcatttggag ttggccaagg tgggcaaagg gttggatatt ctccactgtg
     1081 taatgtttcc tgagccttta tacggtctac ctttgtttgg ctgtgacatt gtggccggcc
     1141 ccggtggagt aagtgcggct attgcggatc tatcccccac ccaaagcgat cgccaattgc
     1201 ccgcagcgta ccaaaaatca ttggcagagc taggccagcc agaatttgag caacaacggg
     1261 aattgccccc ctggggagaa atattttctg aatattgttt attcatccgt cccagcaatg
     1321 tcactgaaga agaaagattt gtacaaaggg tagtggactt tttgcaaatt cattgtcacc
     1381 aatccatcgt tgccgaaccc ttgtctgaag ctcaaacttt ggagcaccgt caggggcaaa
     1441 ttcattactg ccaacaacaa cagaaaaatg ataaaacccg tcgggtactg gaaaaagctt
     1501 ttggggaagc ttgggcggaa cggtatatga gccaagtctt atttgatgtt atccaataat
     1561 ctagaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     1621 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
     1681 tttctgcgtt tataggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttt
     1741 ttacggctag ctcagtccta ggtatagtgc tagctccgcc gctatcacat aaggctctaa
     1801 taccatggcc accaccgtac aactcagcga ccaatccctc cgtcagctag aaaccctcgc
     1861 catccacacc gcccacctga ttcagcccca cggtttagtg gtggtcctgc aggaaccaga
     1921 cctcaccatc agccaaatta gcgccaactg caccggcatt ttagggcgat cgccagagga
     1981 tttgttgggc agaaccctag gggaagtgtt tgatagcttt cagattgatc ccatccagag
     2041 tcgcctaacg gccggacaaa tcagcagcct caaccccagt aaactttggg cgcgggtcat
     2101 gggggacgac tttgtcattt ttgacggggt ttttcatcgc aacagtgacg gtttattggt
     2161 atgtgaactc gagccagcct acacttccga taatctgccc ttcctcggtt tttatcacat
     2221 ggccaacgct gccctgaatc ggttgcgcca acaagctaat ctacgggatt tctacgatgt
     2281 tattgtcgaa gaagtccgcc gtatgactgg ctttgaccgg gtgatgctat accgctttga
     2341 tgaaaataac cacggtgatg tcattgccga agataaacgg gatgatatgg aaccctattt
     2401 gggcctgcac tatcccgaat cggatattcc ccaacccgcc cgtcggctat ttatccacaa
     2461 ccccattcga gtaattcccg atgtttatgg tgtggcggtg cccctgaccc cagcggttaa
     2521 ccccagcacc aaccgagcgg tggatttaac agaatccatt ctgcgcagtg cgtaccattg
     2581 ccacttgacc tatctgaaaa atatgggggt aggagcgtct ttaaccattt ccctaattaa
     2641 ggacggccat ctctgggggc tcattgcctg ccaccatcaa acccccaaag taattccctt
     2701 tgaactgcgt aaagcctgcg aattttttgg tcgggtggtg tttagcaaca tttccgccca
     2761 ggaagatacg gaaaccttcg attaccgggt gcagctggcg gagcatgaag cggttttatt
     2821 ggacaaaatg accacggcgg cggattttgt cgaaggatta actaatcatc ccgatcgcct
     2881 gttgggatta acgggctccc agggggcggc catttgcttt ggggaaaaat tgattttagt
     2941 aggggaaacc ccggacgaga aagcagtgca atatttactg caatggttgg agaatcggga
     3001 agtgcaagac gttttcttca cctcttccct ctcacaaatt tatcctgatg cagtgaattt
     3061 taaatccgtg gccagtggct tattggccat tcccattgcc cgtcacaact ttttgctctg
     3121 gtttcgccct gaagtgttgc aaacggttaa ttggggcggt gacccaaatc atgcttacga
     3181 agctacccag gaagacggta aaatcgagct ccatccccgc caatcctttg acctctggaa
     3241 agaaattgtc cgactccaat ctttgccctg gcaatcggtg gaaatccaaa gtgccctggc
     3301 cctgaaaaag gcgatcgtca acctcatttt gcgccaggca gaagaattgc atatggcggc
     3361 tggtgttaag caactggcgg atgaccgcac gctgctgatg gcgggggtaa gtcacgactt
     3421 gcgcacgccg ctgacgcgta ttcgcctggc gactgagatg atgagcgagc aggatggcta
     3481 tctggcagaa tcgatcaata aagatatcga agagtgcaac gccatcattg agcagtttat
     3541 cgactacctg cgcaccgggc aggagatgcc gatggaaatg gcggatctta atgcagtact
     3601 cggtgaggtg attgctgccg aaagtggcta tgagcgggaa attgaaaccg cgctttaccc
     3661 cggcagcatt gaagtgaaaa tgcacccgct gtcgatcaaa cgcgcggtgg cgaatatggt
     3721 ggtcaacgcc gcccgttatg gcaatggctg gatcaaagtc agcagcggaa cggagccgaa
     3781 tcgcgcctgg ttccaggtgg aagatgacgg tccgggaatt gcgccggaac aacgtaagca
     3841 cctgttccag ccgtttgtcc gcggcgacag tgcgcgcacc attagcggca cgggattagg
     3901 gctggcaatt gtgcagcgta tcgtggataa ccataacggg atgctggagc ttggcaccag
     3961 cgagcgggtc gggctttcca ttcgcgcctg gctgccagtg ccggtaacgc gggcgcaggg
     4021 cacgacaaaa gaagggtaat ctagaggcat caaataaaac gaaaggctca gtcgaaagac
     4081 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     4141 ccgccctaga ggatccagca agcgaaccgg aattgccagc tggggcgccc tctggtaagg
     4201 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
     4261 ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgagg gaagcggtga
     4321 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     4381 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     4441 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     4501 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     4561 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     4621 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     4681 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     4741 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     4801 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     4861 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     4921 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     4981 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     5041 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataagagc
     5101 tcgcttggac tcctgttgat agatccagta atgacctcag aactccatct ggatttgttc
     5161 agaacgctcg gttgccgccg ggcgtttttt attggtgaga atccaagcac tagtaacaac
     5221 ttatatcgta tggggctgac ttcaggtgct acatttgaag agataaattg cactgaaatc
     5281 tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg tctgcgcgta
     5341 atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga
     5401 gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc aaaacttgtc
     5461 ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa atcaattacc
     5521 agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa gacgatagtt
     5581 accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt ccagcttgga
     5641 gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc ggccataaca
     5701 gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg gagccgccag
     5761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga tttgagcgtc
     5821 agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt gccgcggccc
     5881 tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc cccgttcgta
     5941 agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc
     6001 ggaatatatc ccctaggtgg agtctggcct caaatacaaa gtctgcaaag tctcgaaata
     6061 ttaaaccaga gaataaataa ttaaacgcca aaatatttct agcaaaaacc ccagttatta
     6121 aaccgcctaa gtcccccagg aaagggggat ataacagtat agattttgtc agccttcagc
     6181 ttggctttac cgtcaaaaaa a
//
