LOCUS       62056_10375        5203 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 5203)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             72..116
                     /codon_start=1
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /translation="KETAAAKFERQHMDS"
     terminator      169..216
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      260..715
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(833..1690)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1691..1783)
                     /label=AmpR promoter
     rep_origin      1864..2452
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(2638..2780)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2885..3073)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(3611..3632)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3648..4727)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4728..4805)
                     /label=lacI promoter
     promoter        5121..5139
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5140..5164
                     /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             5179..5201
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tacatatggc agatctcaat tggatatcgg ccggccacgc gatcgctgac gtcggtaccc
       61 tcgagtctgg taaagaaacc gctgctgcga aatttgaacg ccagcacatg gactcgtcta
      121 ctagtcgcag cttaattaac ctaaactgct gccaccgctg agcaataact agcataaccc
      181 cttggggcct ctaaacgggt cttgaggggt tttttgctag cgaaaggagg agtcgactat
      241 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      301 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      361 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      421 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      481 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      541 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      601 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      661 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttctggc
      721 ggcacgatgg catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
      781 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
      841 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      901 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
      961 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1021 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1081 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1141 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1201 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1261 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     1321 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     1381 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     1441 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     1501 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     1561 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     1621 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     1681 gaatactcat actcttcctt tttcaatcat gattgaagca tttatcaggg ttattgtctc
     1741 atgagcggat acatatttga atgtatttag aaaaataaac aaataggtca tgaccaaaat
     1801 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1861 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1921 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1981 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2041 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2101 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2161 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2221 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2281 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2341 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2401 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2461 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2521 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2581 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2641 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatat atggtgcact
     2701 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     2761 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     2821 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     2881 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     2941 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3001 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3061 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3121 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3181 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3241 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     3301 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     3361 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     3421 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     3481 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg ctagtcatgc
     3541 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
     3601 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
     3661 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3721 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     3781 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     3841 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     3901 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
     3961 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     4021 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     4081 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     4141 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     4201 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     4261 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     4321 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     4381 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     4441 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     4501 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     4561 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     4621 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     4681 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     4741 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     4801 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag
     4861 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc
     4921 caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag
     4981 cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac
     5041 cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagccta ggatcgagat
     5101 cgatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     5161 ttcccctcta gaaataattt tgtttaactt taagaaggag ata
//
