LOCUS       62056_9770        8702 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 8702)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8702
                     /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)"
     CDS             661..1377
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3913..3930
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     terminator      3997..4044
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4081..4536
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4562..4666
                     /label=AmpR promoter
     CDS             4667..5524
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5698..6286
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(6472..6611)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(6716..6904)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(7679..7700)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(join(7716..8702,1..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 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      721 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      781 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      841 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      901 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      961 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1021 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1081 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     1141 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1201 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1261 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1321 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagaag
     1381 cttaacggtg ataccgggca gtttgttcgc ggggatgccg tacgcacaac catcaataaa
     1441 aacggtcgcc agattgtgag agctgaagga acggcaaata ccactgtggt ttatgccggc
     1501 ggcgaccaga ctgtacatgg tcacgcactg gataccacgc tgaatggggg ataccagtat
     1561 gtgcacaacg gcggtacagc gtctgacact gttgtgaaca gtgacggctg gcagattgtc
     1621 aaaaacgggg gtgtggccgg gaataccacc gttaatcaga agggcagact gcaggtggac
     1681 gccggtggta cagccacgaa tgtcaccctg aagcagggcg gcgcactggt taccagtacg
     1741 gctgcaaccg ttaccggcat aaaccgcctg ggagcattct ctgttgtgga gggtaaagct
     1801 gataatgtcg tactggaaaa tggcggacgc ctggatgtgc tgaccggaca cacagccact
     1861 aatacccgcg tggatgatgg cggaacgctg gatgtccgca acggtggcac cgccaccacc
     1921 gtatccatgg gaaatggcgg tgtactgctg gccgattccg gtgccgctgt cagtggtacc
     1981 cggagcgacg gaaaggcatt cagtatcgga ggcggtcagg cggatgccct gatgctggaa
     2041 aaaggcagtt cattcacgct gaacgccggt gatacggcca cggataccac ggtaaatggc
     2101 ggactgttca ccgccagggg cggcacactg gcgggcacca ccacgctgaa taacggcgcc
     2161 atacttaccc tttccgggaa gacggtgaac aacgataccc tgaccatccg tgaaggcgat
     2221 gcactcctgc agggaggctc tctcaccggt aacggcagcg tggaaaaatc aggaagtggc
     2281 acactcactg tcagcaacac cacactcacc cagaaagccg tcaacctgaa tgaaggcacg
     2341 ctgacgctga acgacagtac cgtcaccacg gatgtcattg ctcagcgcgg tacagccctg
     2401 aagctgaccg gcagcactgt gctgaacggt gccattgacc ccacgaatgt cactctcgcc
     2461 tccggtgcca cctggaatat ccccgataac gccacggtgc agtcggtggt ggatgacctc
     2521 agccatgccg gacagattca tttcacctcc acccgcacag ggaagttcgt accggcaacc
     2581 ctgaaagtga aaaacctgaa cggacagaat ggcaccatca gcctgcgtgt acgcccggat
     2641 atggcacaga acaatgctga cagactggtc attgacggcg gcagggcaac cggaaaaacc
     2701 atcctgaacc tggtgaacgc cggcaacagt gcgtcggggc tggcgaccag cggtaagggt
     2761 attcaggtgg tggaagccat taacggtgcc accacggagg aaggggcctt tgtccagggg
     2821 aacaggctgc aggccggtgc ctttaactac tccctcaacc gggacagtga tgagagctgg
     2881 tatctgcgca gtgaaaatgc ttatcgtgca gaagtccccc tgtatgcctc catgctgaca
     2941 caggcaatgg actatgaccg gattgtggca ggctcccgca gccatcagac cggtgtaaat
     3001 ggtgaaaaca acagcgtccg tctcagcatt cagggcggtc atctcggtca cgataacaat
     3061 ggcggtattg cccgtggggc cacgccggaa agcagcggca gctatggatt cgtccgtctg
     3121 gagggtgacc tgatgagaac agaggttgcc ggtatgtctg tgaccgcggg ggtatatggt
     3181 gctgctggcc attcttccgt tgatgttaag gatgatgacg gctcccgtgc cggcacggtc
     3241 cgggatgatg ccggcagcct gggcggatac ctgaatctgg tacacacgtc ctccggcctg
     3301 tgggctgaca ttgtggcaca gggaacccgc cacagcatga aagcgtcatc ggacaataac
     3361 gacttccgcg cccggggctg gggctggctg ggctcactgg aaaccggtct gcccttcagt
     3421 atcactgaca acctgatgct ggagccacaa ctgcagtata cctggcaggg actttccctg
     3481 gatgacggta aggacaacgc cggttatgtg aagttcgggc atggcagtgc acaacatgtg
     3541 cgtgccggtt tccgtctggg cagccacaac gatatgacct ttggcgaagg cacctcatcc
     3601 cgtgcccccc tgcgtgacag tgcaaaacac agtgtgagtg aattaccggt gaactggtgg
     3661 gtacagcctt ctgttatccg caccttcagc tcccggggag atatgcgtgt ggggacttcc
     3721 actgcaggca gcgggatgac gttctctccc tcacagaatg gcacatcact ggacctgcag
     3781 gccggactgg aagcccgtgt ccgggaaaat atcaccctgg gcgttcaggc cggttatgcc
     3841 cacagcgtca gcggcagcag cgctgaaggg tataacggtc aggccacact gaatgtgacc
     3901 ttctgactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
     3961 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     4021 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
     4081 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4141 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4201 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4261 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4321 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4381 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4441 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4501 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4561 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4621 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4681 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4741 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4801 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4861 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4921 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4981 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5041 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5101 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5161 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     5221 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5281 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5341 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5401 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5461 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5521 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5581 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5641 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5701 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5761 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5821 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5881 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5941 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6001 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6061 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6121 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6181 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6241 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6361 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6421 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     6481 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca
     6541 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     6601 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     6661 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     6721 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     6781 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     6841 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     6901 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     6961 gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt
     7021 gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc
     7081 aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg
     7141 cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg
     7201 aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc
     7261 agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc
     7321 gccagcctag ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca
     7381 tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg
     7441 cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc
     7501 tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga
     7561 gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc
     7621 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     7681 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7741 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7801 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     7861 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     7921 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     7981 cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     8041 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     8101 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     8161 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     8221 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     8281 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     8341 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     8401 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     8461 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     8521 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     8581 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     8641 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     8701 gc
//
