LOCUS       62056_14355        7533 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 7533)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7533
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        247..277
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    285..301
                     /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)."
     primer_bind     309..325
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     sig_peptide     371..433
                     /label=OmpA signal peptide
                     /note="signal peptide from the E. coli outer membrane
                     protein OmpA"
     CDS             458..475
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     promoter        1840..1917
                     /label=lacI promoter
     CDS             1918..2997
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     protein_bind    3013..3034
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             5099..5287
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    5392..5532
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(5718..6306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6480..7337)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7338..7442)
                     /label=AmpR promoter
ORIGIN
        1 ggataaccag aagcaataaa aaatcaaatc ggatttcact atataatctc actttatcta
       61 agatgaatcc gatggaagca tcctgttttc tctcaatttt tttatctaaa acccagcgtt
      121 cgatgcttct ttgagcgaac gatcaaaaat aagtgccttc ccatcaaaaa aatattctca
      181 acataaaaaa ctttgtgtaa tacttgtaac gctacatgga gattaactca atctagctag
      241 agaggcttta cactttatgc ttccggctcg tataatgtgt ggaattgtga gcggataaca
      301 atttcacaca ggaaacagct atgaccatga ttacggattc actggaactc tagataacga
      361 ggcgcaaaaa atgaaaaaga cagctatcgc gattgcagtg gcactggcag gtttcgctac
      421 cgtcgctcag gctggatccg cagaattcag cgctagccac catcatcatc atcattaagc
      481 ttagatccgg ctgagcaacg acgtgaacgc aatgcgttcc gacgttcagg ctgctaaaga
      541 tgacgcagct cgtgctaacc agcgtctgga caacatggct actaaatacc gcaagtaata
      601 gtacctgtga agtgaaaaat ggcgcacatt gtgcgacatt ttttttgtct gccgtttacc
      661 gctactgcgt cacgcgtaac atattccctt gctctggttc accattctgc gctgactcta
      721 ctgaaggcgc attgctggct gcgggagttg ctccactgct caccgaaacc ggataccctg
      781 cccgacgata caacgcttta tcgactaact tctgatctac agccttattg tctttaaatt
      841 gcgtaaagcc tgctggcagt gtgtatggca ttgtctgaac gttctgctgt tcttctgccg
      901 atagtggtcg atgtacttca acataacgca tcccgttagg ctccacggaa tatttcaccg
      961 gttcgttgat cactttcacc ggcgttcccg tccgcacgct ggagaacaag gctttaatat
     1021 ccggtgcatt catgcgaata caccctgaac tgacgcgcaa accgacgctg tccggcgcac
     1081 tggtaccatg aatgaggtat tcgccattac catgcgcgag gcgcagtgcg taacgtccta
     1141 gcgggttatt tggtccggca ggaacgactg gcggtaattt aatgccacgc tccagcgaac
     1201 gctgacgaat gcctgccgta ggcgtccagg ttgggttagg gattttctgc ccaacacgcg
     1261 tttccatcac cggcgtttcc agcccctgca atccaatacc tattggataa acctgcacaa
     1321 tattttctcc cggcggataa taataaaggc gcagctctgc aaggttggtc gacggtcgac
     1381 cggtctcact tcaacataac gcatcccgtt aggctccacg gaatatttca ccggttcgtt
     1441 gatcactttc accggcgttc ccgtccgcac gctggagaac aaggctttaa tatccggtgc
     1501 attcatgcga atacaccctg aactgacgcg caaaccgacg ctgtccggcg cactggtacc
     1561 atgaatgagg tattcgccat taccatgcgc gaggcgcagt gcgtaacgtc ctagcgggtt
     1621 atttggtccg gcaggaacga ctggcggtaa tttaatgcca cgctccagcg aacgctgacg
     1681 aatgcctgcc gtaggcgtcc aggttgggtt agggattttc tgcccaacac gcgtttccat
     1741 caccggcgtt tccagcccct gcaatccaat acctattgga taaacctgca caatattttc
     1801 tcccggcgga taataataaa ggcgcagctc tgcaaggttg acaccatcga atggcgcaaa
     1861 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
     1921 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     1981 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     2041 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     2101 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     2161 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     2221 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     2281 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     2341 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     2401 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     2461 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
     2521 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
     2581 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
     2641 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
     2701 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
     2761 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     2821 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
     2881 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
     2941 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     3001 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     3061 gctaacgata atccctgacg cggtgcatca ggtaataaca gttgtgaagg aatagttatc
     3121 gtcgtaccag gttttggcac cggggcgata gtgttattgg cttcaaggat caacattgcc
     3181 gcagtatcaa aacgtcgggc aatagcctga aggtttttat ccccttcttg caccgtatac
     3241 gtttgatttt gcccaaccag tcggcttccg gttggtggta gcggataatc aaccgcccag
     3301 gcagcctgga tggcgctaaa agcgccgata agcgtgagtg taagcaaaga cgcgcgtttc
     3361 attgtaaacc tcctgtattt gccggagact cacgctgaaa cgtcggatgg cgcttatgtt
     3421 cacctgaaac caaaacactc ctgtgcaggt cagtgtaaac attgaccatc cggcaatgtg
     3481 agccaaccgg atgaaagctg tccttttagt ttagctaagt gcagcggctt tggcgcgaat
     3541 tgcgcgaatc atcgcttcca gaccttgtga acgagatggg gtgagatgtt gggtgagcgc
     3601 cattttttca aaccacggac gcacatcgaa attgacaata tcctgcggcg tcatctgatc
     3661 gtagagaata aagacgaccg caataagccc tttcacaatc gccgcatcgc tgtcgccctg
     3721 taattcaata attccctggg cattctggcg catgacaatc cacacctgac tctgacagcc
     3781 ctgaatgcta ttttgtggac ttctgtcttc gtcgcgtaat tctggcagac gctgggggac
     3841 cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta
     3901 tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag
     3961 cgctctgggt cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt
     4021 cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca
     4081 ccaaacgttt cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct
     4141 acgtcttgct ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg
     4201 cttccggcgg catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg
     4261 accatcaggg acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcactg
     4321 gaccgctgat cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat
     4381 ggattgtagg cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga
     4441 gccgggccac ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc
     4501 aagaattgga gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca
     4561 gaacatatcc atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggcagcgt
     4621 tgggtcctgg ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg
     4681 gcggggttgc cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga
     4741 ctgctgctgc aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt
     4801 ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat
     4861 cgcaggatgc tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca
     4921 ttgaccctga gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc
     4981 acaacgttcc agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc
     5041 tcgtttcatc ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt
     5101 gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac
     5161 gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct
     5221 tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga
     5281 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     5341 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
     5401 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
     5461 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
     5521 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5581 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5641 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5701 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5761 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5821 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5881 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5941 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6001 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6061 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6121 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     6181 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6241 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6301 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6361 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6421 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6481 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6541 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6601 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6661 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6721 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6781 gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6841 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6901 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6961 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7021 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7081 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7141 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7201 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7261 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7321 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7381 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7441 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     7501 acctataaaa ataggcgtat cacgaggccc ttt
//
