LOCUS       62056_13000        3558 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 3558)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        618..636
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             668..690
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             709..726
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             736..747
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     terminator      874..921
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(1287..1315)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     primer_bind     complement(1337..1353)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1361..1377)
                     /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(1385..1415)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1430..1451)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1739..2327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2501..3358)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3359..3463)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc aaggagatgg
      421 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
      481 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
      541 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
      601 tctcgatccc gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag
      661 aaataatttt gtttaacttt aagaaggaga tataccatgt ctggttctca tcatcatcat
      721 catcatagca gcggcatcga aggccgcggc cgcttaatta aaaccatggc aaagtcgact
      781 cgagcgagct ctgcagcccg ggatccgtaa gatccggctg ctaacaaagc ccgaaaggaa
      841 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
      901 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatatcc acaggacggg
      961 tgtggtcgcc atgatcgcgt agtcgatagt ggctccaagt agcgaagcga gcaggactgg
     1021 gcggcggcca aagcggtcgg acagtgctcc gagaacgggt gcgcatagaa attgcatcaa
     1081 cgcatatagc gctagcagca cgccatagtg actggcgatg ctgtcggaat ggacgatatc
     1141 ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac
     1201 ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta
     1261 gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg
     1321 agaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1381 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1441 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1501 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     1921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     1981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2161 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2461 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2521 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2581 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2641 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2701 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2761 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2821 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     2881 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     2941 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3001 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3061 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3121 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3181 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3241 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3301 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3481 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3541 tcacgaggcc ctttcgtc
//
