LOCUS       62056_8185        6398 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 6398)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..54
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    62..78
                     /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             1475..1564
                     /codon_start=1
                     /label=Twin-Strep-tag
                     /note="two Strep-Tag(R)II moieties connected by a linker 
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     promoter        complement(1564..1592)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     rep_origin      1733..2188
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2215..2319
                     /label=AmpR promoter
     CDS             2320..3177
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3351..3939
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4125..4267)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4676..4849)
                     /codon_start=1
                     /label=lacZ-alpha
                     /note="LacZ-alpha fragment of beta-galactosidase"
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     protein_bind    complement(4869..4885)
                     /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)."
     promoter        complement(4893..4923)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4938..4959)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(4975..6054)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(6055..6132)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     protein_bind    6184..6205
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6220..6250
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
ORIGIN
        1 gatcagctta ctccccatcc ccctgttgac aattaatcat cggctcgtat aatgtgtgga
       61 attgtgagcg gataacaatt tcacacagga aacaggatcc atggccaaga agaccagcag
      121 caagggcaag ctgcccccct acattttcag ccccattccc ttcctgggcc acgccattgc
      181 ttttggaaaa agccccatcg agttcctgga gaacgcctac gagaagtacg gccccgtgtt
      241 tagctttacc atggtgggca agaccttcac ctacctgctg ggcagcgatg ccgctgctct
      301 gctgtttaat agcaaaaatg aggacctgaa cgccgaggac gtgtacagca gactgaccac
      361 ccctgtgttc ggcaagggcg tggcttatga tgtgcccaat cctgtgttcc tggaacagaa
      421 aaagatgctg aagagcggcc tgaacatcgc ccacttcaag cagcacgtga gcatcatcga
      481 gaaggaaacc aaggagtact tcgagagctg gggcgagagc ggcgaaaaga atgtgtttga
      541 ggccctgagc gagctgatca tcctgaccgc tagccactgc ctgcatggca aagagatcag
      601 aagccagctg aacgagaagg tggcccagct gtatgccgac ctggatggcg gatttagcca
      661 tgctgcttgg ctgctgcctg gctggctgcc tctgcctagc tttagaagaa gagacagagc
      721 ccacagagag atcaaggaca tcttctacaa ggccatccag aagagaagac agagccagga
      781 gaagatcgac gacatcctgc agaccctgct ggacgccaca tataaggacg gaagacccct
      841 gacagacgac gaggtggctg gaatgctgat tggactgctg ctggctggac agcacacaag
      901 cagcacaaca agcgcttgga tgggcttctt cctggccaga gacaagaccc tgcagaagaa
      961 gtgctacctg gagcagaaga ccgtgtgcgg cgaaaacctg cctcctctga catatgacca
     1021 actgaaggac ctgaacctgc tggacagatg catcaaggaa accctgagac tcagaccccc
     1081 catcatgatc atgatgagaa tggccagaac cccccagacc gtggccggat atacaatccc
     1141 ccctggacat caagtgtgtg tgagccccac agtgaaccaa agactgaagg acagctgggt
     1201 ggagagactg gacttcaacc ccgacagata cctgcaggac aaccccgcca gcggagaaaa
     1261 atttgcttac gtgcctttcg gcgccggaag acacagatgc attggcgaga atttcgccta
     1321 cgtgcagatc aagaccatct ggagcaccat gctgagactg tacgagttcg acctgatcga
     1381 cggctacttc cccaccgtga attacaccac catgatccac acccccgaga accccgtgat
     1441 tagatacaag agaagaagca agcaccacca ccacagcgcc tggagccacc cgcagttcga
     1501 aaaaggcggc ggcagcggcg gcggcagcgg cggcagcgcc tggagccacc cgcagttcga
     1561 aaaataaaag cttatcgatg ataagctgtc aaacatgagc agatctgagc ccgcctaatg
     1621 agcgggcttt tttttcagat ctgcttgaag acgaaagggc ctcgtgatac gcctattttt
     1681 ataggttaat gtcatgataa taatggtttc ttagcgtcaa agcaaccata gtacgcgccc
     1741 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     1801 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     1861 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     1921 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     1981 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2041 ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt
     2101 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2161 tttaacaaaa tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga
     2221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2521 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     2581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     2881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     3001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     3061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     3121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     3181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     3241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     3301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3481 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     3541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     4021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     4081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     4141 ctccttacgc atctgtgcgg tatttcacac cgcatatatg gtgcactctc agtacaatct
     4201 gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat
     4261 ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     4321 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     4381 accgtcatca ccgaaacgcg cgaggcagaa cgccatcaaa aataattcgc gtctggcctt
     4441 cctgtagcca gctttcatca acattaaatg tgagcgagta acaacccgtc ggattctccg
     4501 tgggaacaaa cggcggattg accgtaatgg gataggttac gttggtgtag atgggcgcat
     4561 cgtaaccgtg catctgccag tttgagggga cgacgacagt atcggcctca ggaagatcgc
     4621 actccagcca gctttccggc accgcttctg gtgccggaaa ccaggcaaag cgccattcgc
     4681 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4741 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     4801 agtcacgacg ttgtaaaacg acggccagtg aatccgtaat catggtcata gctgtttcct
     4861 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4921 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4981 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     5041 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     5101 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     5161 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     5221 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
     5281 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     5341 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     5401 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     5461 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     5521 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     5581 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     5641 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     5701 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     5761 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     5821 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     5881 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     5941 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     6001 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     6061 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcacca
     6121 ttcgatggtg tcctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     6181 aattaatgta agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     6241 tcgtataatg tgtggaattg tgagcggata acaatttcac acaggaaaca ggatcgatcc
     6301 atcgatgagc ttactcccca tccccctgtt gacaattaat catcggctcg tataatgtgt
     6361 ggaattgtga gcggataaca atttcacaca ggaaacag
//
