LOCUS       62056_8605        9770 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9770)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1079..1095
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             1314..1336
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1344..2057
                     /codon_start=1
                     /label=GFPuv
                     /note="GFP variant optimized for excitation by UV light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2109..2125)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2133..2149)
                     /label=lac operator
                     /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        complement(2157..2187)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2202..2223)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2890..3738)
                     /codon_start=1
                     /label=RSF1010 RepC
                     /note="replication protein C of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="VVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
                     GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
                     TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
                     YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
                     TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
                     AG"
     CDS             complement(3728..4564)
                     /codon_start=1
                     /label=RSF1010 RepA
                     /note="replication protein A of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
                     LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
                     GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
                     EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
                     WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
     CDS             complement(5078..6046)
                     /codon_start=1
                     /label=RSF1010 RepB
                     /note="replication protein B of the broad-host-range
                     plasmid RSF1010 (Scholz et al., 1989)"
                     /translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
                     QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
                     YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
                     PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
                     EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
                     YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
     oriT            complement(7285..7372)
                     /direction=LEFT
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      7714..8108
                     /label=RSF1010 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     RSF1010; requires the RSF1010 RepA/B/C proteins for 
                     replication (Scholz et al., 1989)"
     CDS             complement(8925..9737)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tatcgcgagc ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc
       61 tcgagcaaga cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt
      121 aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga
      181 gattttgaga cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc
      241 agatcacgca tcttcccgac aacgcagacc gttccgtggc aaagcaaaag ttcaaaatca
      301 ccaactggtc cacctacaac aaagctctca tcaaccgtgg ctccctcact ttctggctgg
      361 atgatggggc gattcaggcc tggtatgagt cagcaacacc ttcttcacga ggcagacctc
      421 agcgctattc tgaccttgcc atcacgactg tgctggtcat taaacgcgta ttcaggctga
      481 ccctgcgcgc tgcgcagggc tttattgatt ccatttttac actgatgaat gttccgttgc
      541 gctgcccgga ttacagctgc gtttcctgtt tggggtcgtt tgcgggaagg ggcggaatcc
      601 tacgctaagg ctttggccag cgatattctc cggtgagatt gatgtgttcc caggggatag
      661 gagaagtcgc ttgatatcta gtatgacgtc tgtcgcacct gcttgatcgc ggccgcgata
      721 gctagatcgc gttgctcctc ttctccatcc gcgttccaag ctgcggaaag gcacccataa
      781 gcgtacgcct ggtcgagcag gcgacgcgga tcgacgtcca gcgcacgaga gaatgcgtcc
      841 gccatctgtg caatgcgtct aggatcgaga caaaggtcgt ctctgtcagc cggatcgtag
      901 aacatattgg cggcgccaaa gcccacttca ccgaccagac attcgccatt caggctgcgc
      961 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     1021 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     1081 aaaacgacgg ccagtgaatt cgagctcggt acccggggat ctagaggatc tcaatgaata
     1141 ttggttgaca cgggcgtata agacatgtta tactgttgaa taacaagctt tcaattctct
     1201 attttgagaa gaattaatgt gcttgggagt ccctgatgat taaattaaat aaaccaagat
     1261 tttaccatga ctgcaatttt agagagacgg gggatcgatc ctctagaaat aattttgttt
     1321 aactttaaga aggagatata catatgagta aaggagaaga acttttcact ggagttgtcc
     1381 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
     1441 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
     1501 tacctgttcc atggccaaca cttgtcacta ctttctctta tggtgttcaa tgcttttccc
     1561 gttatccgga tcatatgaaa cggcatgact ttttcaagag tgccatgccc gaaggttatg
     1621 tacaggaacg cactatatct ttcaaagatg acgggaacta caagacgcgt gctgaagtca
     1681 agtttgaagg tgataccctt gttaatcgta tcgagttaaa aggtattgat tttaaagaag
     1741 atggaaacat tctcggacac aaactcgagt acaactataa ctcacacaat gtatacatca
     1801 cggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattcgccac aacattgaag
     1861 atggatcagt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     1921 tccttttacc agacaaccat tacctgtcga cacaatctgc cctttcgaaa gatcccaacg
     1981 aaaagcgtga ccacatggtc cttcttgagt ttgtaactgc tgctgggatt acacatggca
     2041 tggatgagct ctacaaataa ggatcctcta gagtcgacct gcaggcatgc aagcttggcg
     2101 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     2161 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     2221 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     2281 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggccacg cttcccgaag
     2341 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2401 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2461 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2521 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2581 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2641 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa agaagagcat
     2701 acatctggaa gcaaagccag gaaagcggcc tatggagctg tgcggcagcg ctcagtaggc
     2761 aatttttcaa aatattgtta agccttttct gagcatggta tttttcatgg tattaccaat
     2821 tagcaggaaa ataagccatt gaatataaaa gataaaaatg tcttgtttac aatagagtgg
     2881 ggggggtcag cctgccgcct tgggccgggt gatgtcgtac ttgcccgccg cgaactcggt
     2941 taccgtccag cccagcgcga ccagctccgg caacgcctcg cgcacccgct ggcggcgctt
     3001 gcgcatggtc gaaccactgg cctctgacgg ccagacatag ccgcacaagg tatctatgga
     3061 agccttgccg gttttgccgg ggtcgatcca gccacacagc cgctggtgca gcaggcgggc
     3121 ggtttcgctg tccagcgccc gcacctcgtc catgctgatg cgcacatgct ggccgccacc
     3181 catgacggcc tgcgcgatca aggggttcag ggccacgtac aggcgcccgt ccgcctcgtc
     3241 gctggcgtac tccgacagca gccgaaaccc ctgccgcttg cggccattct gggcgatgat
     3301 ggataccttc caaaggcgct cgatgcagtc ctgtatgtgc ttgagcgccc caccactatc
     3361 gacctctgcc ccgatttcct ttgccagcgc ccgatagcta cctttgacca ccatggcatc
     3421 agcggtgacg gcctcccact tgggttccag gaacagccgg agctgccgtc cgccttcggt
     3481 cttgggttcc gggccaagca ctaggccatt aggcccagcc atggccacca gcccttgcag
     3541 gatgcgcaga tcatcagcgc ccagcggctc cgggccgctg aactcgatcc gcttgccgtc
     3601 gccgtagtca tacgtcacgt ccagcttgct gcgcttgcgc tcgccccgct tgagggcacg
     3661 gaacaggccg ggggccagac agtgcgccgg gtcgtgccgg acgtggctga ggctgtgctt
     3721 gttcttaggc ttcaccacgg ggcaccccct tgctcttgcg ctgcctctcc agcacggcgg
     3781 gcttgagcac cccgccgtca tgccgcctga accaccgatc agcgaacggt gcgccatagt
     3841 tggccttgct cacaccgaag cggacgaaga accggcgctg gtcgtcgtcc acaccccatt
     3901 cctcggcctc ggcgctggtc atgctcgaca ggtaggactg ccagcggatg ttatcgacca
     3961 gtaccgagct gccccggctg gcctgctgct ggtcgcctgc gcccatcatg gccgcgccct
     4021 tgctggcatg gtgcaggaac acgatagagc acccggtatc ggcggcgatg gcctccatgc
     4081 gaccgatgac ctgggccatg gggccgctgg cgttttcttc ctcgatgtgg aaccggcgca
     4141 gcgtgtccag caccatcagg cggcggccct cggcggcgcg cttgaggccg tcgaaccact
     4201 ccggggccat gatgttgggc aggctgccga tcagcggctg gatcagcagg ccgtcagcca
     4261 cggcttgccg ttcctcggcg ctgaggtgcg ccccaagggc gtgcaggcgg tgatgaatgg
     4321 cggtgggcgg gtcttcggcg ggcaggtaga tcaccgggcc ggtgggcagt tcgcccacct
     4381 ccagcagatc cggcccgcct gcaatctgtg cggccagttg cagggccagc atggatttac
     4441 cggcaccacc gggcgacacc agcgccccga ccgtaccggc caccatgttg ggcaaaacgt
     4501 agtccagcgg tggcggcgct gctgcgaacg cctccagaat attgataggc ttatgggtag
     4561 ccattgattg cctcctttgc aggcagttgg tggttaggcg ctggcggggt cactaccccc
     4621 gccctgcgcc gctctgagtt cttccaggca ctcgcgcagc gcctcgtatt cgtcgtcggt
     4681 cagccagaac ttgcgctgac gcatcccttt ggccttcatg cgctcggcat atcgcgcttg
     4741 gcgtacagcg tcagggctgg ccagcaggtc gccggtctgc ttgtcctttt ggtctttcat
     4801 atcagtcacc gagaaacttg ccggggccga aaggcttgtc ttcgcggaac aaggacaagg
     4861 tgcagccgtc aaggttaagg ctggccatat cagcgactga aaagcggcca gcctcggcct
     4921 tgtttgacgt ataaccaaag ccaccgggca accaatagcc cttgtcactt ttgatcaggt
     4981 agaccgaccc tgaagcgctt ttttcgtatt ccataaaacc cccttctgtg cgtgagtact
     5041 catagtataa caggcgtgag taccaacgca agcactacat gctgaaatct ggcccgcccc
     5101 tgtccatgcc tcgctggcgg ggtgccggtg cccgtgccag ctcggcccgc gcaagctgga
     5161 cgctgggcag acccatgacc ttgctgacgg tgcgctcgat gtaatccgct tcgtggccgg
     5221 gcttgcgctc tgccagcgct gggctggcct cggccatggc cttgccgatt tcctcggcac
     5281 tgcggccccg gctggccagc ttctgcgcgg cgataaagtc gcacttgctg aggtcatcac
     5341 cgaagcgctt gaccagcccg gccatctcgc tgcggtactc gtccagcgcc gtgcgccggt
     5401 ggcggctaag ctgccgctcg ggcagttcga ggctggccag cctgcgggcc ttctcctgct
     5461 gccgctgggc ctgctcgatc tgctggccag cctgctgcac cagcgccggg ccagcggtgg
     5521 cggtcttgcc cttggattca cgcagcagca cccacggctg ataaccggcg cgggtggtgt
     5581 gcttgtcctt gcggttggtg aagcccgcca agcggccata gtggcggctg tcggcgctgg
     5641 ccgggtcggc gtcgtactcg ctggccagcg tccgggcaat ctgcccccga agttcaccgc
     5701 ctgcggcgtc ggccaccttg acccatgcct gatagttctt cgggctggtt tccactacca
     5761 gggcaggctc ccggccctcg gctttcatgt catccaggtc aaactcgctg aggtcgtcca
     5821 ccagcaccag accatgccgc tcctgctcgg cgggcctgat atacacgtca ttgccctggg
     5881 cattcatccg cttgagccat ggcgtgttct ggagcacttc ggcggctgac cattcccggt
     5941 tcatcatctg gccggtggtg gcgtccctga cgccgatatc gaagcgctca cagcccatgg
     6001 ccttgagctg tcggcctatg gcctgcaaag tcctgtcgtt cttcatcggg ccaccaagcg
     6061 cagccagatc gagccgtcct cggttgtcag tggcgtcagg tcgagcaaga gcaacgatgc
     6121 gatcagcagc accaccgtag gcatcatgga agccagcatc acggttagcc atagcttcca
     6181 gtgccacccc cgcgacgcgc tccgggcgct ctgcgcggcg ctgctcacct cggcggctac
     6241 ctcccgcaac tctttggcca gctccaccca tgccgcccct gtctggcgct gggctttcag
     6301 ccactccgcc gcctgcgcct cgctggcctg ctgggtctgg ctcatgacct gccgggcttc
     6361 gtcggccagt gtcgccatgc tctgggccag cggttcgatc tgctccgcta actcgttgat
     6421 gcctctggat ttcttcactc tgtcgattgc gttcatggtc tattgcctcc cggtattcct
     6481 gtaagtcgat gatctgggcg ttggcggtgt cgatgttcag ggccacgtct gcccggtcgg
     6541 tgcggatgcc ccggccttcc atctccacca cgttcggccc caggtgaaca ccgggcaggc
     6601 gctcgatgcc ctgcgcctca agtgttctgt ggtcaatgcg ggcgtcgtgg ccagcccgct
     6661 ctaatgcccg gttggcatgg tcggcccatg cctcgcgggt ctgctcaagc catgccttgg
     6721 gcttgagcgc ttcggtcttc tgtgccccgc ccttctccgg ggtcttgccg ttgtaccgct
     6781 tgaaccactg agcggcgggc cgctcgatgc cgtcattgat ccgctcggag atcatcaggt
     6841 ggcagtgcgg gttctcgccg ccaccggcat ggatggccag cgtatacggc aggcgctcgg
     6901 caccggtcag gtgctgggcg aactcggacg ccagcgcctt ctgctggtcg agggtcagct
     6961 cgaccggcag ggcaaattcg acctccttga acagccgccc attggcgcgt tcatacaggt
     7021 cggcagcatc ccagtagtcg gcgggccgct cgacgaactc cggcatgtgc ccggattcgg
     7081 cgtgcaagac ttcatccatg tcgcgggcat acttgccttc gcgctggatg tagtcggcct
     7141 tggccctggc cgattggccg cccgacctgc tgccggtttt cgccgtaagg tgataaatcg
     7201 ccatgctgcc tcgctgttgc ttttgctttt cggctccatg caatggccct cggagagcgc
     7261 accgcccgaa gggtggccgt taggccagtt tctcgaagag aaaccggtaa gtgcgccctc
     7321 ccctacaaag tagggtcggg attgccgccg ctgtgcctcc atgatagcct acgagacagc
     7381 acattaacaa tggggtgtca agatggttaa ggggagcaac aaggcggcgg atcggctggc
     7441 caagctcgaa gaacaacgag cgcgaatcaa tgccgaaatt cagcgggtgc gggcaaggga
     7501 acagcagcaa gagcgcaaga acgaaacaag gcgcaaggtg ctggtggggg ccatgatttt
     7561 ggccaaggtg aacagcagcg agtggccgga ggatcggctc atggcggcaa tggatgcgta
     7621 ccttgaacgc gaccacgacc gcgccttgtt cggtctgccg ccacgccaga aggatgagcc
     7681 gggctgaatg atcgaccgag acaggccctg cggggctgca cacgcgcccc cacccttcgg
     7741 gtagggggaa aggccgctaa agcggctaaa agcgctccag cgtatttctg cggggtttgg
     7801 tgtggggttt agcgggcttt gcccgccttt ccccctgccg cgcagcggtg gggcggtgtg
     7861 tagcctagcg cagcgaatag accagctatc cggcctctgg ccgggcatat tgggcaaggg
     7921 cagcagcgcc ccacaagggc gctgataacc gcgcctagtg gattattctt agataatcat
     7981 ggatggattt ttccaacacc ccgccagccc ccgcccctgc tgggtttgca ggtttggggg
     8041 cgtgacagtt attgcagggg ttcgtgacag ttattgcagg ggggcgtgac agttattgca
     8101 ggggttcgtg acagttagta cgggagtgac gggcactggc tggcaatgtc tagcaacggc
     8161 aggcatttcg gctgagggta aaagaacttt ccgctaagcg atagactgta tgtaaacaca
     8221 gtattgcaag gacgcggaac atgcctcatg tggcggccag gacggccagc cgggatcggg
     8281 atactggtcg ttaccagagc caccgacccg agcaaaccct tctctatcag atcgttgacg
     8341 agtattaccc ggcattcgct gcgcttatgg cagagcaggg aaaggaattg ccgggctatg
     8401 tgcaacggga atttgaagaa tttctccaat gcgggcggct ggagcatggc tttctacggg
     8461 ttcgctgcga gtcttgccac gccgagcacc tggtcgcttt cagctgtaat ccgggcagcg
     8521 caacggaaca ttcatcagtg taaaaatgga atcaataaag ccctgcgcag cgcgcagggt
     8581 cagcctgaat acgcgtttaa tgaccagcac agtcgtgatg gcaaggtcag aatagcgctg
     8641 aggtctgcct cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc
     8701 cagccagaaa gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt
     8761 gattttgaac ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg
     8821 atccttcaac tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc
     8881 gtaatgctct gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc
     8941 atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc
     9001 cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg
     9061 tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca
     9121 aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc
     9181 aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca
     9241 aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat
     9301 acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac
     9361 actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat
     9421 gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa
     9481 tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct
     9541 gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc
     9601 ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta
     9661 tacccatata aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc
     9721 cgttgaatat ggctcatagc accccttgta ttactgttta tgtaagcaga
//
