LOCUS       62056_18600        4779 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 4779)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4779
                     /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      643..729
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      821..848
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        868..959
                     /label=AmpR promoter
     CDS             960..1817
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1991..2579
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(2827..3255)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3580..3657
                     /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             3658..4737
                     /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 catgggatcc ggaattcaaa
      361 ggcctacgtc gacgagctca ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg
      421 aggcatgcgg taccaagctt ggctgttttg gcggatgaga gaagattttc agcctgatac
      481 agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc ggcagtagcg
      541 cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc gccgatggta
      601 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      661 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      721 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
      781 gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat
      841 ggcctttttg cgtttctaca aactcttttt gtttattttt ctaaatacat tcaaatatgt
      901 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      961 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     1021 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1081 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1141 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1201 gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1261 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1321 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1381 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1441 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1501 ctacagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1561 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1621 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     1681 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     1741 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1801 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1861 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1921 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1981 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     2041 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2101 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     2161 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2221 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2281 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2341 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2401 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2461 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2521 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2581 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2641 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2701 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     2761 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcataattt
     2821 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2881 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2941 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3001 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3061 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3121 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3181 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3241 cgctacaggg cgcgtcccat tcgccattca ggctgctatg gtgcactctc agtacaatct
     3301 gctctgatgc cgcatagtta agccagtacc agtcacgtag cgatatcgga gtgtatacac
     3361 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3421 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3481 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat
     3541 caattcgcgc gcgaaggcga agcggcatgc atttacgttg acaccatcga atggtgcaaa
     3601 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
     3661 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     3721 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     3781 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     3841 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     3901 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     3961 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     4021 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     4081 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     4141 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     4201 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggctgg
     4261 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
     4321 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
     4381 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
     4441 ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
     4501 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
     4561 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
     4621 tcactggtga aaagaaaaac caccctggca cccaatacgc aaaccgcctc tccccgcgcg
     4681 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     4741 gcgcaacgca attaatgtga gttagcgcga attgatctg
//
