LOCUS       62056_10330        7115 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 7115)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7115
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             83..100
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             119..1087
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVGDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDSLDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     CDS             1118..1834
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     promoter        1909..1927
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2061..2105
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      2157..2204
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2241..2696
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2814..3671)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3672..3764)
                     /label=AmpR promoter
     rep_origin      3845..4433
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4619..4761)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4866..5054)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(5592..5613)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5629..6708)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6709..6786)
                     /label=lacI promoter
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattctat
      121 gaagcgtgcg cctgttattc caaaacatac gctcaatact caaccggttg gagatacttc
      181 gttatcgaca ccagctgccc cgatggtgga ttcgttaatt gcgcgcgtag gagtaatggc
      241 tcgcggtaat gccattactt tgcctgtatg tggtcgggat gtgaagttta ctcttgaagt
      301 gctccggggt gatagtgttg agaagacctc tcgggtatgg tcaggtaatg aacgtgacca
      361 ggagctgctt actgaggact cactggatga tctcatccct tcttttctac tgactggtca
      421 acagacaccg gcgttcggtc gaagagtatc tggtgtcata gaaattgccg atgggagtcg
      481 ccgtcgtaaa gctgctgcac ttaccgaaag tgattatcgt gttctggttg gcgagctgga
      541 tgatgagcag atggctgcat tatccagatt gggtaacgat tatcgcccaa caagtgctta
      601 tgaacgtggt cagcgttatg caagccgatt gcagaatgaa tttgctggaa atatttctgc
      661 gctggctgat gcggaaaata tttcacgtaa gattattacc cgctgtatca acacagccaa
      721 attgcctaaa tcagttgttg ctcttttttc tcaccccggt gaactatctg cccggtcagg
      781 tgatgcactt caaaaagcct ttacagataa agaggaatta cttaagcagc aggcatctaa
      841 ccttcatgag cagaaaaaag ctggggtgat atttgaagct gaagaagtta tcactctttt
      901 aacttctgtg cttaaaacgt catctgcatc aagaactagt ttaagctcac gacatcagtt
      961 tgctcctgga gcgacagtat tgtataaggg cgataaaatg gtgcttaacc tggacaggtc
     1021 tcgtgttcca actgagtgta tagagaaaat tgaggccatt cttaaggaac ttgaaaagcc
     1081 agcacccggc ggcggaggga gtggaggagg gggaagcatg gtgagcaagg gcgaggagct
     1141 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1201 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1261 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     1321 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1381 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     1441 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1501 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     1561 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     1621 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1681 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1741 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1801 cgggatcact ctcggcatgg acgagctgta caagtgaaag cttgcggccg cataatgctt
     1861 aagtcgaaca gaaagtaatc gtattgtaca cggccgcata atcgaaatta atacgactca
     1921 ctatagggga attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag
     1981 aaggagatat acatatggca gatctcaatt ggatatcggc cggccacgcg atcgctgacg
     2041 tcggtaccct cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc cagcacatgg
     2101 actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag caataactag
     2161 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     2221 tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2281 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2341 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2401 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2461 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2521 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2581 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2641 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caatttctgg
     2701 cggcacgatg gcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     2761 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     2821 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     2881 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     2941 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3001 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3061 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     3121 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     3181 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     3241 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     3301 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     3361 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     3421 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     3481 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     3541 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     3601 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     3661 tgaatactca tactcttcct ttttcaatca tgattgaagc atttatcagg gttattgtct
     3721 catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggtc atgaccaaaa
     3781 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3841 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3901 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3961 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     4021 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4081 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4141 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4201 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4261 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4321 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4381 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4441 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4501 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4561 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4621 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tatggtgcac
     4681 tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta
     4741 cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4801 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4861 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca
     4921 gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt
     4981 ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt
     5041 tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata
     5101 ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca tgcccggtta
     5161 ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca gagaaaaatc
     5221 actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca gggtagccag
     5281 cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt ccgcgtttcc
     5341 agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt cgcagacgtt
     5401 ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg ctaaccagta
     5461 aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat gctagtcatg
     5521 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
     5581 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
     5641 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5701 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
     5761 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
     5821 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
     5881 ggtatcgtcg tatcccacta ccgagatgtc cgcaccaacg cgcagcccgg actcggtaat
     5941 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
     6001 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
     6061 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
     6121 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
     6181 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
     6241 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
     6301 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
     6361 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
     6421 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
     6481 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
     6541 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
     6601 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
     6661 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
     6721 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
     6781 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
     6841 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
     6901 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6961 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     7021 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg
     7081 atctcgatcc cgcgaaatta atacgactca ctata
//
