LOCUS       62056_17170        8828 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 8828)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8828
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(69..85)
                     /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(93..121)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     promoter        351..428
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             complement(1790..2638)
                     /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(2628..3464)
                     /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(3978..4946)
                     /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(6185..6272)
                     /direction=LEFT
                     /label=RSF1010 oriT
                     /note="origin of transfer of the broad-host-range plasmid 
                     RSF1010 (Scholz et al., 1989)"
     rep_origin      6614..7008
                     /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(7448..8305)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8306..8397)
                     /label=AmpR promoter
     terminator      complement(8416..8443)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(8535..8621)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     misc_feature    complement(join(8824..8828,1..52))
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat tctgtttcct
       61 gtgtgaaatt gttatccgct cacaattcca cacattatac gagccgatga ttaattgtca
      121 acagctcatt tcagaatatt tgccagaacc gttatgatgt cggcgcaaaa aacattatcc
      181 agaacgggag tgcgccttga gcgacacgaa ttatgcagtg atttacgacc tgcacagcca
      241 taccacagct tccgatggct gcctgacgcc agaagcattg gtgcaccgtg cagtcgataa
      301 gcccggatca gcttgcaatt cgcgcgcgaa ggcgaagcgg catttacgtt gacaccatcg
      361 aatggtgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
      421 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
      481 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag
      541 tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg
      601 gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc
      661 aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga
      721 tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac
      781 gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag
      841 ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca
      901 gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg
      961 gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc
     1021 tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag
     1081 gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg
     1141 ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta
     1201 ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac gataccgaag
     1261 acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc
     1321 aaaccacgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     1381 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     1441 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     1501 cgggcagtga gcgcaacgca attaatgtaa gttagcgcga attgcaagct ggccgacgcg
     1561 ctgggctacg tcttgctggc gttcgggagc agaagagcat acatctggaa gcaaagccag
     1621 gaaagcggcc tatggagctg tgcggcagcg ctcagtaggc aatttttcaa aatattgtta
     1681 agccttttct gagcatggta tttttcatgg tattaccaat tagcaggaaa ataagccatt
     1741 gaatataaaa gataaaaatg tcttgtttac aatagagtgg ggggggtcag cctgccgcct
     1801 tgggccgggt gatgtcgtac ttgcccgccg cgaactcggt taccgtccag cccagcgcga
     1861 ccagctccgg caacgcctcg cgcacccgct ggcggcgctt gcgcatggtc gaaccactgg
     1921 cctctgacgg ccagacatag ccgcacaagg tatctatgga agccttgccg gttttgccgg
     1981 ggtcgatcca gccacacagc cgctggtgca gcaggcgggc ggtttcgctg tccagcgccc
     2041 gcacctcgtc catgctgatg cgcacatgct ggccgccacc catgacggcc tgcgcgatca
     2101 aggggttcag ggccacgtac aggcgcccgt ccgcctcgtc gctggcgtac tccgacagca
     2161 gccgaaaccc ctgccgcttg cggccattct gggcgatgat ggataccttc caaaggcgct
     2221 cgatgcagtc ctgtatgtgc ttgagcgccc caccactatc gacctctgcc ccgatttcct
     2281 ttgccagcgc ccgatagcta cctttgacca ccatggcatc agcggtgacg gcctcccact
     2341 tgggttccag gaacagccgg agctgccgtc cgccttcggt cttgggttcc gggccaagca
     2401 ctaggccatt aggcccagcc atggccacca gcccttgcag gatgcgcaga tcatcagcgc
     2461 ccagcggctc cgggccgctg aactcgatcc gcttgccgtc gccgtagtca tacgtcacgt
     2521 ccagcttgct gcgcttgcgc tcgccccgct tgagggcacg gaacaggccg ggggccagac
     2581 agtgcgccgg gtcgtgccgg acgtggctga ggctgtgctt gttcttaggc ttcaccacgg
     2641 ggcaccccct tgctcttgcg ctgcctctcc agcacggcgg gcttgagcac cccgccgtca
     2701 tgccgcctga accaccgatc agcgaacggt gcgccatagt tggccttgct cacaccgaag
     2761 cggacgaaga accggcgctg gtcgtcgtcc acaccccatt cctcggcctc ggcgctggtc
     2821 atgctcgaca ggtaggactg ccagcggatg ttatcgacca gtaccgagct gccccggctg
     2881 gcctgctgct ggtcgcctgc gcccatcatg gccgcgccct tgctggcatg gtgcaggaac
     2941 acgatagagc acccggtatc ggcggcgatg gcctccatgc gaccgatgac ctgggccatg
     3001 gggccgctgg cgttttcttc ctcgatgtgg aaccggcgca gcgtgtccag caccatcagg
     3061 cggcggccct cggcggcgcg cttgaggccg tcgaaccact ccggggccat gatgttgggc
     3121 aggctgccga tcagcggctg gatcagcagg ccgtcagcca cggcttgccg ttcctcggcg
     3181 ctgaggtgcg ccccaagggc gtgcaggcgg tgatgaatgg cggtgggcgg gtcttcggcg
     3241 ggcaggtaga tcaccgggcc ggtgggcagt tcgcccacct ccagcagatc cggcccgcct
     3301 gcaatctgtg cggccagttg cagggccagc atggatttac cggcaccacc gggcgacacc
     3361 agcgccccga ccgtaccggc caccatgttg ggcaaaacgt agtccagcgg tggcggcgct
     3421 gctgcgaacg cctccagaat attgataggc ttatgggtag ccattgattg cctcctttgc
     3481 aggcagttgg tggttaggcg ctggcggggt cactaccccc gccctgcgcc gctctgagtt
     3541 cttccaggca ctcgcgcagc gcctcgtatt cgtcgtcggt cagccagaac ttgcgctgac
     3601 gcatcccttt ggccttcatg cgctcggcat atcgcgcttg gcgtacagcg tcagggctgg
     3661 ccagcaggtc gccggtctgc ttgtcctttt ggtctttcat atcagtcacc gagaaacttg
     3721 ccggggccga aaggcttgtc ttcgcggaac aaggacaagg tgcagccgtc aaggttaagg
     3781 ctggccatat cagcgactga aaagcggcca gcctcggcct tgtttgacgt ataaccaaag
     3841 ccaccgggca accaatagcc cttgtcactt ttgatcaggt agaccgaccc tgaagcgctt
     3901 ttttcgtatt ccataaaacc cccttctgtg cgtgagtact catagtataa caggcgtgag
     3961 taccaacgca agcactacat gctgaaatct ggcccgcccc tgtccatgcc tcgctggcgg
     4021 ggtgccggtg cccgtgccag ctcggcccgc gcaagctgga cgctgggcag acccatgacc
     4081 ttgctgacgg tgcgctcgat gtaatccgct tcgtggccgg gcttgcgctc tgccagcgct
     4141 gggctggcct cggccatggc cttgccgatt tcctcggcac tgcggccccg gctggccagc
     4201 ttctgcgcgg cgataaagtc gcacttgctg aggtcatcac cgaagcgctt gaccagcccg
     4261 gccatctcgc tgcggtactc gtccagcgcc gtgcgccggt ggcggctaag ctgccgctcg
     4321 ggcagttcga ggctggccag cctgcgggcc ttctcctgct gccgctgggc ctgctcgatc
     4381 tgctggccag cctgctgcac cagcgccggg ccagcggtgg cggtcttgcc cttggattca
     4441 cgcagcagca cccacggctg ataaccggcg cgggtggtgt gcttgtcctt gcggttggtg
     4501 aagcccgcca agcggccata gtggcggctg tcggcgctgg ccgggtcggc gtcgtactcg
     4561 ctggccagcg tccgggcaat ctgcccccga agttcaccgc ctgcggcgtc ggccaccttg
     4621 acccatgcct gatagttctt cgggctggtt tccactacca gggcaggctc ccggccctcg
     4681 gctttcatgt catccaggtc aaactcgctg aggtcgtcca ccagcaccag accatgccgc
     4741 tcctgctcgg cgggcctgat atacacgtca ttgccctggg cattcatccg cttgagccat
     4801 ggcgtgttct ggagcacttc ggcggctgac cattcccggt tcatcatctg gccggtggtg
     4861 gcgtccctga cgccgatatc gaagcgctca cagcccatgg ccttgagctg tcggcctatg
     4921 gcctgcaaag tcctgtcgtt cttcatcggg ccaccaagcg cagccagatc gagccgtcct
     4981 cggttgtcag tggcgtcagg tcgagcaaga gcaacgatgc gatcagcagc accaccgtag
     5041 gcatcatgga agccagcatc acggttagcc atagcttcca gtgccacccc cgcgacgcgc
     5101 tccgggcgct ctgcgcggcg ctgctcacct cggcggctac ctcccgcaac tctttggcca
     5161 gctccaccca tgccgcccct gtctggcgct gggctttcag ccactccgcc gcctgcgcct
     5221 cgctggcctg ctgggtctgg ctcatgacct gccgggcttc gtcggccagt gtcgccatgc
     5281 tctgggccag cggttcgatc tgctccgcta actcgttgat gcctctggat ttcttcactc
     5341 tgtcgattgc gttcatggtc tattgcctcc cggtattcct gtaagtcgat gatctgggcg
     5401 ttggcggtgt cgatgttcag ggccacgtct gcccggtcgg tgcggatgcc ccggccttcc
     5461 atctccacca cgttcggccc caggtgaaca ccgggcaggc gctcgatgcc ctgcgcctca
     5521 agtgttctgt ggtcaatgcg ggcgtcgtgg ccagcccgct ctaatgcccg gttggcatgg
     5581 tcggcccatg cctcgcgggt ctgctcaagc catgccttgg gcttgagcgc ttcggtcttc
     5641 tgtgccccgc ccttctccgg ggtcttgccg ttgtaccgct tgaaccactg agcggcgggc
     5701 cgctcgatgc cgtcattgat ccgctcggag atcatcaggt ggcagtgcgg gttctcgccg
     5761 ccaccggcat ggatggccag cgtatacggc aggcgctcgg caccggtcag gtgctgggcg
     5821 aactcggacg ccagcgcctt ctgctggtcg agggtcagct cgaccggcag ggcaaattcg
     5881 acctccttga acagccgccc attggcgcgt tcatacaggt cggcagcatc ccagtagtcg
     5941 gcgggccgct cgacgaactc cggcatgtgc ccggattcgg cgtgcaagac ttcatccatg
     6001 tcgcgggcat acttgccttc gcgctggatg tagtcggcct tggccctggc cgattggccg
     6061 cccgacctgc tgccggtttt cgccgtaagg tgataaatcg ccatgctgcc tcgctgttgc
     6121 ttttgctttt cggctccatg caatggccct cggagagcgc accgcccgaa gggtggccgt
     6181 taggccagtt tctcgaagag aaaccggtaa gtgcgccctc ccctacaaag tagggtcggg
     6241 attgccgccg ctgtgcctcc atgatagcct acgagacagc acattaacaa tggggtgtca
     6301 agatggttaa ggggagcaac aaggcggcgg atcggctggc caagctcgaa gaacaacgag
     6361 cgcgaatcaa tgccgaaatt cagcgggtgc gggcaaggga acagcagcaa gagcgcaaga
     6421 acgaaacaag gcgcaaggtg ctggtggggg ccatgatttt ggccaaggtg aacagcagcg
     6481 agtggccgga ggatcggctc atggcggcaa tggatgcgta ccttgaacgc gaccacgacc
     6541 gcgccttgtt cggtctgccg ccacgccaga aggatgagcc gggctgaatg atcgaccgag
     6601 acaggccctg cggggctgca cacgcgcccc cacccttcgg gtagggggaa aggccgctaa
     6661 agcggctaaa agcgctccag cgtatttctg cggggtttgg tgtggggttt agcgggcttt
     6721 gcccgccttt ccccctgccg cgcagcggtg gggcggtgtg tagcctagcg cagcgaatag
     6781 accagctatc cggcctctgg ccgggcatat tgggcaaggg cagcagcgcc ccacaagggc
     6841 gctgataacc gcgcctagtg gattattctt agataatcat ggatggattt ttccaacacc
     6901 ccgccagccc ccgcccctgc tgggtttgca ggtttggggg cgtgacagtt attgcagggg
     6961 ttcgtgacag ttattgcagg ggggcgtgac agttattgca ggggttcgtg acagttagta
     7021 cgggagtgac gggcactggc tggcaatgtc tagcaacggc aggcatttcg gctgagggta
     7081 aaagaacttt ccgctaagcg atagactgta tgtaaacaca gtattgcaag gacgcggaac
     7141 atgcctcatg tggcggccag gacggccagc cgggatcggg atactggtcg ttaccagagc
     7201 caccgacccg agcaaaccct tctctatcag atcgttgacg agtattaccc ggcattcgct
     7261 gcgcttatgg cagagcaggg aaaggaattg ccgggctatg tgcaacggga atttgaagaa
     7321 tttctccaat gcgggcggct ggagcatggc tttctacggg ttcgctgcga gtcttgccac
     7381 gccgagcacc tggtcgcttt cagaaatcaa tctaaagtat atatgagtaa acttggtctg
     7441 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7501 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7561 gccccagtgc tgcaatgata ccgcgggacc cacgctcacc ggctccagat ttatcagcaa
     7621 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7681 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7741 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7801 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7861 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7921 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7981 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     8041 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     8101 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     8161 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     8221 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     8281 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     8341 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaaaga
     8401 gtttgtagaa acgcaaaaag gccatccgtc aggatggcct tctgcttaat ttgatgcctg
     8461 gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc gttgcttcgc aacgttcaaa
     8521 tccgctcccg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     8581 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc agttccctac
     8641 tctcgcatgg ggagacccca cactaccatc ggcgctacgg cgtttcactt ctgagttcgg
     8701 catggggtca ggtgggacca ccgcgctact gccgccaggc aaattctgtt ttatcagacc
     8761 gcttctgcgt tctgatttaa tctgtatcag gctgaaaatc ttctctcatc cgccaaaaca
     8821 gccaagct
//
