LOCUS       62056_21355        5549 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 5549)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    230..246
                     /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)."
     CDS             279..296
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             318..338
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     terminator      1440..1526
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      1618..1645
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        1665..1756
                     /label=AmpR promoter
     CDS             1757..2614
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2788..3376
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(3624..4052)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4350..4427
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4428..5507
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gtccgtataa tctgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgt cgtactacca tcaccatcac catcacgatt
      301 acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catggtgacg caatctgcaa
      361 tgtgcatccc gctgtggccc gcccggaacg gcaatactgc gcatctggtc atgtgccctt
      421 tcgctggcgg cagcagtagc gcgtttcgcc actggcaagc tgagcaactg actgattgcg
      481 cgctttctct ggtgacctgg ccggggcgcg atcgccttcg ccatctggaa ccgctcagaa
      541 gcattacaca actggcggca ctgctggcga acgagctgga agcatccgta tcgcctgaca
      601 cgccgctttt actcgccggg cacagcatgg gggcgcaggt ggcgtttgaa acctgccgac
      661 ttctggagca acgggggctt gcgccacaag gactgattat ttccgggtgc catgccccgc
      721 atctgcattc tgaacgccag ctcagccatc gcgatgatgc cgactttatc gctgagctga
      781 tagacattgg cggatgttct cctgaactgc gggaaaacca ggaattaatg tcgctgtttc
      841 ttcctcttct gcgcgctgat ttttacgcca ccgagagcta tcactacgac tcgcccgacg
      901 tctgtccgcc gctgcgcacg cctgcgctgt tattgtgcgg cagccacgat cgcgaagcct
      961 cctggcagca ggtcgatgcc tggcgtcagt ggctgagcca cgttacaggc ccggtggtga
     1021 ttgacggcga tcatttctat cccattcaac aagcccggtc cttttttacg cagattgtcc
     1081 gccattttcc ccacgcattt tctgcaatga ccgcgttgca aaaacagccc agtacttcag
     1141 aaaggtgaga attcaaaggc ctacgtcgac gagctcaact agtgcggccg ctttcgaatc
     1201 tagagcctgc agtctcgagg catgcggtac caagcttggc tgttttggcg gatgagagaa
     1261 gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa aacagaattt
     1321 gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag aagtgaaacg
     1381 ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc
     1441 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1501 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     1561 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     1621 aggccatcct gacggatggc ctttttgcgt ttctacaaac tctttttgtt tatttttcta
     1681 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1741 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1801 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1861 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1921 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1981 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2041 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2101 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     2161 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     2221 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     2281 gcgtgacacc acgatgccta cagcaatggc aacaacgttg cgcaaactat taactggcga
     2341 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     2401 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     2461 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     2521 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2581 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2641 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2701 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2761 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2821 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2881 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2941 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3001 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3061 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3121 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     3181 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     3241 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     3301 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     3361 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     3421 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     3481 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3541 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3601 ttcacaccgc ataattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3661 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3721 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3781 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3841 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3901 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3961 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4021 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgctatggtg
     4081 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     4141 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     4201 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     4261 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc
     4321 gcgaaggcga agcggcatgc atttacgttg acaccatcga atggtgcaaa acctttcgcg
     4381 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     4441 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     4501 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     4561 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     4621 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     4681 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     4741 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     4801 actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg
     4861 cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag
     4921 acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt
     4981 tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc
     5041 tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg
     5101 gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg
     5161 ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg
     5221 gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc
     5281 cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc
     5341 tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga
     5401 aaagaaaaac caccctggca cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     5461 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     5521 attaatgtga gttagcgcga attgatctg
//
