LOCUS       62056_8610        9731 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 9731)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9731
                     /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..2018
                     /codon_start=1
                     /label=DsRed2
                     /note="improved tetrameric variant of DsRed fluorescent
                     protein"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(2070..2086)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2094..2110)
                     /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(2118..2148)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2163..2184)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2851..3699)
                     /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(3689..4525)
                     /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(5039..6007)
                     /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(7246..7333)
                     /direction=LEFT
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      7675..8069
                     /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(8886..9698)
                     /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 catatggcct cctccgagaa cgtcatcacc gagttcatgc
     1381 gcttcaaggt gcgcatggag ggcaccgtga acggccacga gttcgagatc gagggcgagg
     1441 gcgagggccg cccctacgag ggccacaaca ccgtgaagct gaaggtgacc aagggcggcc
     1501 ccctgccctt cgcctgggac atcctgtccc cccagttcca gtacggctcc aaggtgtacg
     1561 tgaagcaccc cgccgacatc cccgactaca agaagctgtc cttccccgag ggcttcaagt
     1621 gggagcgcgt gatgaacttc gaggacggcg gcgtggcgac cgtgacccag gactcctccc
     1681 tgcaggacgg ctgcttcatc tacaaggtga agttcatcgg cgtgaacttc ccctccgacg
     1741 gccccgtgat gcagaagaag accatgggct gggaggcctc caccgagcgc ctgtaccccc
     1801 gcgacggcgt gctgaagggc gagacccaca aggccctgaa gctgaaggac ggcggccact
     1861 acctggtgga gttcaagtcc atctacatgg ccaagaagcc cgtgcagctg cccggctact
     1921 actacgtgga cgccaagctg gacatcacct cccacaacga ggactacacc atcgtggagc
     1981 agtacgagcg caccgagggc cgccaccacc tgttcctgta gggatcctct agagtcgacc
     2041 tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     2101 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     2161 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     2221 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2281 attgggccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2341 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2401 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2461 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2521 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2581 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2641 aggaagcgga aagaagagca tacatctgga agcaaagcca ggaaagcggc ctatggagct
     2701 gtgcggcagc gctcagtagg caatttttca aaatattgtt aagccttttc tgagcatggt
     2761 atttttcatg gtattaccaa ttagcaggaa aataagccat tgaatataaa agataaaaat
     2821 gtcttgttta caatagagtg gggggggtca gcctgccgcc ttgggccggg tgatgtcgta
     2881 cttgcccgcc gcgaactcgg ttaccgtcca gcccagcgcg accagctccg gcaacgcctc
     2941 gcgcacccgc tggcggcgct tgcgcatggt cgaaccactg gcctctgacg gccagacata
     3001 gccgcacaag gtatctatgg aagccttgcc ggttttgccg gggtcgatcc agccacacag
     3061 ccgctggtgc agcaggcggg cggtttcgct gtccagcgcc cgcacctcgt ccatgctgat
     3121 gcgcacatgc tggccgccac ccatgacggc ctgcgcgatc aaggggttca gggccacgta
     3181 caggcgcccg tccgcctcgt cgctggcgta ctccgacagc agccgaaacc cctgccgctt
     3241 gcggccattc tgggcgatga tggatacctt ccaaaggcgc tcgatgcagt cctgtatgtg
     3301 cttgagcgcc ccaccactat cgacctctgc cccgatttcc tttgccagcg cccgatagct
     3361 acctttgacc accatggcat cagcggtgac ggcctcccac ttgggttcca ggaacagccg
     3421 gagctgccgt ccgccttcgg tcttgggttc cgggccaagc actaggccat taggcccagc
     3481 catggccacc agcccttgca ggatgcgcag atcatcagcg cccagcggct ccgggccgct
     3541 gaactcgatc cgcttgccgt cgccgtagtc atacgtcacg tccagcttgc tgcgcttgcg
     3601 ctcgccccgc ttgagggcac ggaacaggcc gggggccaga cagtgcgccg ggtcgtgccg
     3661 gacgtggctg aggctgtgct tgttcttagg cttcaccacg gggcaccccc ttgctcttgc
     3721 gctgcctctc cagcacggcg ggcttgagca ccccgccgtc atgccgcctg aaccaccgat
     3781 cagcgaacgg tgcgccatag ttggccttgc tcacaccgaa gcggacgaag aaccggcgct
     3841 ggtcgtcgtc cacaccccat tcctcggcct cggcgctggt catgctcgac aggtaggact
     3901 gccagcggat gttatcgacc agtaccgagc tgccccggct ggcctgctgc tggtcgcctg
     3961 cgcccatcat ggccgcgccc ttgctggcat ggtgcaggaa cacgatagag cacccggtat
     4021 cggcggcgat ggcctccatg cgaccgatga cctgggccat ggggccgctg gcgttttctt
     4081 cctcgatgtg gaaccggcgc agcgtgtcca gcaccatcag gcggcggccc tcggcggcgc
     4141 gcttgaggcc gtcgaaccac tccggggcca tgatgttggg caggctgccg atcagcggct
     4201 ggatcagcag gccgtcagcc acggcttgcc gttcctcggc gctgaggtgc gccccaaggg
     4261 cgtgcaggcg gtgatgaatg gcggtgggcg ggtcttcggc gggcaggtag atcaccgggc
     4321 cggtgggcag ttcgcccacc tccagcagat ccggcccgcc tgcaatctgt gcggccagtt
     4381 gcagggccag catggattta ccggcaccac cgggcgacac cagcgccccg accgtaccgg
     4441 ccaccatgtt gggcaaaacg tagtccagcg gtggcggcgc tgctgcgaac gcctccagaa
     4501 tattgatagg cttatgggta gccattgatt gcctcctttg caggcagttg gtggttaggc
     4561 gctggcgggg tcactacccc cgccctgcgc cgctctgagt tcttccaggc actcgcgcag
     4621 cgcctcgtat tcgtcgtcgg tcagccagaa cttgcgctga cgcatccctt tggccttcat
     4681 gcgctcggca tatcgcgctt ggcgtacagc gtcagggctg gccagcaggt cgccggtctg
     4741 cttgtccttt tggtctttca tatcagtcac cgagaaactt gccggggccg aaaggcttgt
     4801 cttcgcggaa caaggacaag gtgcagccgt caaggttaag gctggccata tcagcgactg
     4861 aaaagcggcc agcctcggcc ttgtttgacg tataaccaaa gccaccgggc aaccaatagc
     4921 ccttgtcact tttgatcagg tagaccgacc ctgaagcgct tttttcgtat tccataaaac
     4981 ccccttctgt gcgtgagtac tcatagtata acaggcgtga gtaccaacgc aagcactaca
     5041 tgctgaaatc tggcccgccc ctgtccatgc ctcgctggcg gggtgccggt gcccgtgcca
     5101 gctcggcccg cgcaagctgg acgctgggca gacccatgac cttgctgacg gtgcgctcga
     5161 tgtaatccgc ttcgtggccg ggcttgcgct ctgccagcgc tgggctggcc tcggccatgg
     5221 ccttgccgat ttcctcggca ctgcggcccc ggctggccag cttctgcgcg gcgataaagt
     5281 cgcacttgct gaggtcatca ccgaagcgct tgaccagccc ggccatctcg ctgcggtact
     5341 cgtccagcgc cgtgcgccgg tggcggctaa gctgccgctc gggcagttcg aggctggcca
     5401 gcctgcgggc cttctcctgc tgccgctggg cctgctcgat ctgctggcca gcctgctgca
     5461 ccagcgccgg gccagcggtg gcggtcttgc ccttggattc acgcagcagc acccacggct
     5521 gataaccggc gcgggtggtg tgcttgtcct tgcggttggt gaagcccgcc aagcggccat
     5581 agtggcggct gtcggcgctg gccgggtcgg cgtcgtactc gctggccagc gtccgggcaa
     5641 tctgcccccg aagttcaccg cctgcggcgt cggccacctt gacccatgcc tgatagttct
     5701 tcgggctggt ttccactacc agggcaggct cccggccctc ggctttcatg tcatccaggt
     5761 caaactcgct gaggtcgtcc accagcacca gaccatgccg ctcctgctcg gcgggcctga
     5821 tatacacgtc attgccctgg gcattcatcc gcttgagcca tggcgtgttc tggagcactt
     5881 cggcggctga ccattcccgg ttcatcatct ggccggtggt ggcgtccctg acgccgatat
     5941 cgaagcgctc acagcccatg gccttgagct gtcggcctat ggcctgcaaa gtcctgtcgt
     6001 tcttcatcgg gccaccaagc gcagccagat cgagccgtcc tcggttgtca gtggcgtcag
     6061 gtcgagcaag agcaacgatg cgatcagcag caccaccgta ggcatcatgg aagccagcat
     6121 cacggttagc catagcttcc agtgccaccc ccgcgacgcg ctccgggcgc tctgcgcggc
     6181 gctgctcacc tcggcggcta cctcccgcaa ctctttggcc agctccaccc atgccgcccc
     6241 tgtctggcgc tgggctttca gccactccgc cgcctgcgcc tcgctggcct gctgggtctg
     6301 gctcatgacc tgccgggctt cgtcggccag tgtcgccatg ctctgggcca gcggttcgat
     6361 ctgctccgct aactcgttga tgcctctgga tttcttcact ctgtcgattg cgttcatggt
     6421 ctattgcctc ccggtattcc tgtaagtcga tgatctgggc gttggcggtg tcgatgttca
     6481 gggccacgtc tgcccggtcg gtgcggatgc cccggccttc catctccacc acgttcggcc
     6541 ccaggtgaac accgggcagg cgctcgatgc cctgcgcctc aagtgttctg tggtcaatgc
     6601 gggcgtcgtg gccagcccgc tctaatgccc ggttggcatg gtcggcccat gcctcgcggg
     6661 tctgctcaag ccatgccttg ggcttgagcg cttcggtctt ctgtgccccg cccttctccg
     6721 gggtcttgcc gttgtaccgc ttgaaccact gagcggcggg ccgctcgatg ccgtcattga
     6781 tccgctcgga gatcatcagg tggcagtgcg ggttctcgcc gccaccggca tggatggcca
     6841 gcgtatacgg caggcgctcg gcaccggtca ggtgctgggc gaactcggac gccagcgcct
     6901 tctgctggtc gagggtcagc tcgaccggca gggcaaattc gacctccttg aacagccgcc
     6961 cattggcgcg ttcatacagg tcggcagcat cccagtagtc ggcgggccgc tcgacgaact
     7021 ccggcatgtg cccggattcg gcgtgcaaga cttcatccat gtcgcgggca tacttgcctt
     7081 cgcgctggat gtagtcggcc ttggccctgg ccgattggcc gcccgacctg ctgccggttt
     7141 tcgccgtaag gtgataaatc gccatgctgc ctcgctgttg cttttgcttt tcggctccat
     7201 gcaatggccc tcggagagcg caccgcccga agggtggccg ttaggccagt ttctcgaaga
     7261 gaaaccggta agtgcgccct cccctacaaa gtagggtcgg gattgccgcc gctgtgcctc
     7321 catgatagcc tacgagacag cacattaaca atggggtgtc aagatggtta aggggagcaa
     7381 caaggcggcg gatcggctgg ccaagctcga agaacaacga gcgcgaatca atgccgaaat
     7441 tcagcgggtg cgggcaaggg aacagcagca agagcgcaag aacgaaacaa ggcgcaaggt
     7501 gctggtgggg gccatgattt tggccaaggt gaacagcagc gagtggccgg aggatcggct
     7561 catggcggca atggatgcgt accttgaacg cgaccacgac cgcgccttgt tcggtctgcc
     7621 gccacgccag aaggatgagc cgggctgaat gatcgaccga gacaggccct gcggggctgc
     7681 acacgcgccc ccacccttcg ggtaggggga aaggccgcta aagcggctaa aagcgctcca
     7741 gcgtatttct gcggggtttg gtgtggggtt tagcgggctt tgcccgcctt tccccctgcc
     7801 gcgcagcggt ggggcggtgt gtagcctagc gcagcgaata gaccagctat ccggcctctg
     7861 gccgggcata ttgggcaagg gcagcagcgc cccacaaggg cgctgataac cgcgcctagt
     7921 ggattattct tagataatca tggatggatt tttccaacac cccgccagcc cccgcccctg
     7981 ctgggtttgc aggtttgggg gcgtgacagt tattgcaggg gttcgtgaca gttattgcag
     8041 gggggcgtga cagttattgc aggggttcgt gacagttagt acgggagtga cgggcactgg
     8101 ctggcaatgt ctagcaacgg caggcatttc ggctgagggt aaaagaactt tccgctaagc
     8161 gatagactgt atgtaaacac agtattgcaa ggacgcggaa catgcctcat gtggcggcca
     8221 ggacggccag ccgggatcgg gatactggtc gttaccagag ccaccgaccc gagcaaaccc
     8281 ttctctatca gatcgttgac gagtattacc cggcattcgc tgcgcttatg gcagagcagg
     8341 gaaaggaatt gccgggctat gtgcaacggg aatttgaaga atttctccaa tgcgggcggc
     8401 tggagcatgg ctttctacgg gttcgctgcg agtcttgcca cgccgagcac ctggtcgctt
     8461 tcagctgtaa tccgggcagc gcaacggaac attcatcagt gtaaaaatgg aatcaataaa
     8521 gccctgcgca gcgcgcaggg tcagcctgaa tacgcgttta atgaccagca cagtcgtgat
     8581 ggcaaggtca gaatagcgct gaggtctgcc tcgtgaagaa ggtgttgctg actcatacca
     8641 ggcctgaatc gccccatcat ccagccagaa agtgagggag ccacggttga tgagagcttt
     8701 gttgtaggtg gaccagttgg tgattttgaa cttttgcttt gccacggaac ggtctgcgtt
     8761 gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag
     8821 ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct
     8881 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
     8941 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
     9001 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
     9061 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
     9121 actgaatccg gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc
     9181 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
     9241 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
     9301 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
     9361 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
     9421 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
     9481 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
     9541 aacaactctg gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg
     9601 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
     9661 ggcctcgagc aagacgtttc ccgttgaata tggctcatag caccccttgt attactgttt
     9721 atgtaagcag a
//
