LOCUS       62056_9760        8003 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 8003)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(94..171)
                     /label=lacI promoter
     promoter        480..498
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    499..523
                     /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)."
     RBS             538..560
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     sig_peptide     568..633
                     /label=pelB signal sequence
                     /note="leader peptide for secretion"
     CDS             3214..3231
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3298..3345
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3382..3837
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3863..3967
                     /label=AmpR promoter
     CDS             3968..4825
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4999..5587
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5773..5912)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6017..6205)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(6980..7001)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(join(7017..8003,1..93))
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
       61 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
      121 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
      181 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
      241 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
      301 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
      361 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
      421 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
      481 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
      541 gtttaacttt aagaaggaga tatacatatg aaatacctgc tgccgaccgc tgctgctggt
      601 ctgctgctcc tcgctgccca gccggcgatg gccatggata tcggaattaa ttcggatccg
      661 aattcgagct ccgtcgacaa gcttaacggt gataccgggc agtttgttcg cggggatgcc
      721 gtacgcacaa ccatcaataa aaacggtcgc cagattgtga gagctgaagg aacggcaaat
      781 accactgtgg tttatgccgg cggcgaccag actgtacatg gtcacgcact ggataccacg
      841 ctgaatgggg gataccagta tgtgcacaac ggcggtacag cgtctgacac tgttgtgaac
      901 agtgacggct ggcagattgt caaaaacggg ggtgtggccg ggaataccac cgttaatcag
      961 aagggcagac tgcaggtgga cgccggtggt acagccacga atgtcaccct gaagcagggc
     1021 ggcgcactgg ttaccagtac ggctgcaacc gttaccggca taaaccgcct gggagcattc
     1081 tctgttgtgg agggtaaagc tgataatgtc gtactggaaa atggcggacg cctggatgtg
     1141 ctgaccggac acacagccac taatacccgc gtggatgatg gcggaacgct ggatgtccgc
     1201 aacggtggca ccgccaccac cgtatccatg ggaaatggcg gtgtactgct ggccgattcc
     1261 ggtgccgctg tcagtggtac ccggagcgac ggaaaggcat tcagtatcgg aggcggtcag
     1321 gcggatgccc tgatgctgga aaaaggcagt tcattcacgc tgaacgccgg tgatacggcc
     1381 acggatacca cggtaaatgg cggactgttc accgccaggg gcggcacact ggcgggcacc
     1441 accacgctga ataacggcgc catacttacc ctttccggga agacggtgaa caacgatacc
     1501 ctgaccatcc gtgaaggcga tgcactcctg cagggaggct ctctcaccgg taacggcagc
     1561 gtggaaaaat caggaagtgg cacactcact gtcagcaaca ccacactcac ccagaaagcc
     1621 gtcaacctga atgaaggcac gctgacgctg aacgacagta ccgtcaccac ggatgtcatt
     1681 gctcagcgcg gtacagccct gaagctgacc ggcagcactg tgctgaacgg tgccattgac
     1741 cccacgaatg tcactctcgc ctccggtgcc acctggaata tccccgataa cgccacggtg
     1801 cagtcggtgg tggatgacct cagccatgcc ggacagattc atttcacctc cacccgcaca
     1861 gggaagttcg taccggcaac cctgaaagtg aaaaacctga acggacagaa tggcaccatc
     1921 agcctgcgtg tacgcccgga tatggcacag aacaatgctg acagactggt cattgacggc
     1981 ggcagggcaa ccggaaaaac catcctgaac ctggtgaacg ccggcaacag tgcgtcgggg
     2041 ctggcgacca gcggtaaggg tattcaggtg gtggaagcca ttaacggtgc caccacggag
     2101 gaaggggcct ttgtccaggg gaacaggctg caggccggtg cctttaacta ctccctcaac
     2161 cgggacagtg atgagagctg gtatctgcgc agtgaaaatg cttatcgtgc agaagtcccc
     2221 ctgtatgcct ccatgctgac acaggcaatg gactatgacc ggattgtggc aggctcccgc
     2281 agccatcaga ccggtgtaaa tggtgaaaac aacagcgtcc gtctcagcat tcagggcggt
     2341 catctcggtc acgataacaa tggcggtatt gcccgtgggg ccacgccgga aagcagcggc
     2401 agctatggat tcgtccgtct ggagggtgac ctgatgagaa cagaggttgc cggtatgtct
     2461 gtgaccgcgg gggtatatgg tgctgctggc cattcttccg ttgatgttaa ggatgatgac
     2521 ggctcccgtg ccggcacggt ccgggatgat gccggcagcc tgggcggata cctgaatctg
     2581 gtacacacgt cctccggcct gtgggctgac attgtggcac agggaacccg ccacagcatg
     2641 aaagcgtcat cggacaataa cgacttccgc gcccggggct ggggctggct gggctcactg
     2701 gaaaccggtc tgcccttcag tatcactgac aacctgatgc tggagccaca actgcagtat
     2761 acctggcagg gactttccct ggatgacggt aaggacaacg ccggttatgt gaagttcggg
     2821 catggcagtg cacaacatgt gcgtgccggt ttccgtctgg gcagccacaa cgatatgacc
     2881 tttggcgaag gcacctcatc ccgtgccccc ctgcgtgaca gtgcaaaaca cagtgtgagt
     2941 gaattaccgg tgaactggtg ggtacagcct tctgttatcc gcaccttcag ctcccgggga
     3001 gatatgcgtg tggggacttc cactgcaggc agcgggatga cgttctctcc ctcacagaat
     3061 ggcacatcac tggacctgca ggccggactg gaagcccgtg tccgggaaaa tatcaccctg
     3121 ggcgttcagg ccggttatgc ccacagcgtc agcggcagca gcgctgaagg gtataacggt
     3181 caggccacac tgaatgtgac cttctgactc gagcaccacc accaccacca ctgagatccg
     3241 gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta
     3301 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     3361 atatccggat tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3421 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3481 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3541 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3601 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3661 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3721 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3781 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
     3841 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3901 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3961 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4021 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4081 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4141 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4201 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4261 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4321 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4381 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4441 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4501 cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4561 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4621 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4681 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4741 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4801 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4861 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4921 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     4981 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5041 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5101 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     5161 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5221 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5281 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5341 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5401 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5461 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5521 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5581 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5641 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5701 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5761 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     5821 caatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     5881 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     5941 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6001 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     6061 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     6121 gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa
     6181 gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg
     6241 gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac
     6301 atgcccggtt actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc
     6361 agagaaaaat cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac
     6421 agggtagcca gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact
     6481 tccgcgtttc cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg
     6541 tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct
     6601 gctaaccagt aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca
     6661 tgcgcacccg tggggccgcc atgccggcga taatggcctg cttctcgccg aaacgtttgg
     6721 tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa gattccgaat accgcaagcg
     6781 acaggccgat catcgtcgcg ctccagcgaa agcggtcctc gccgaaaatg acccagagcg
     6841 ctgccggcac ctgtcctacg agttgcatga taaagaagac agtcataagt gcggcgacga
     6901 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     6961 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     7021 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7081 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     7141 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     7201 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     7261 ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg cagcccggac
     7321 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     7381 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     7441 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     7501 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     7561 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     7621 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     7681 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     7741 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     7801 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     7861 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     7921 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     7981 tccacttttt cccgcgtttt cgc
//
