LOCUS       40924_40962        4701 bp DNA     circular SYN 18-DEC-2018
DEFINITION  ORF selection and expression vector pSOS-X, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4701)
  AUTHORS   Cox JC, Lape J, Sayed MA, Hellinga HW.
  TITLE     Protein fabrication automation
  JOURNAL   Protein Sci. 16 (3), 379-390 (2007)
  PUBMED    17242375
REFERENCE   2  (bases 1 to 4701)
  AUTHORS   Cox JC, Hellinga HW.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2006) Biochemistry, Duke University Medical 
            Center, Research Drive, DUMC 3711, Nanaline 413, Durham, NC 27710, 
            USA
REFERENCE   3  (bases 1 to 4701)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4701)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Sci."; date: "2007"; volume: "16"; issue: "3"; pages: "379-390"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (28-SEP-2006) Biochemistry, Duke University Medical Center, Research
            Drive, DUMC 3711, Nanaline 413, Durham, NC 27710, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(240..828)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1002..1859)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1860..1964)
                     /label=AmpR promoter
     regulatory      2106..2151
                     /gene="lacI"
                     /regulatory_class="promoter"
     CDS             2172..3251
                     /label=lacI
                     /note="lac repressor"
     protein_bind    3267..3288
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3503..3531
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3539..3555
                     /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)."
     regulatory      3611..3616
                     /label=Shine-Delgarno sequence
                     /note="Shine-Delgarno sequence"
                     /regulatory_class="ribosome_binding_site"
     misc_feature    3617..3624
                     /label=PacI restriction site
                     /note="PacI restriction site"
     misc_feature    3633..3638
                     /label=NaeI restriction site
                     /note="NaeI restriction site"
     CDS             3639..3656
                     /label=6xHis
                     /note="6xHis affinity tag"
     misc_feature    3657..3659
                     /label=amber stop codon
                     /note="amber stop codon"
     CDS             3666..4319
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     terminator      4492..4578
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4670..4697
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
       61 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      121 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      181 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
      241 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
      301 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
      361 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
      421 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
      481 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
      541 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
      601 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      661 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
      721 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      781 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      841 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      901 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      961 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     1021 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     1081 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     1141 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     1201 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     1261 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctgcaggc
     1321 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     1381 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     1441 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     1501 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     1561 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaacacgg
     1621 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     1681 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     1741 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     1801 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     1861 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     1921 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     1981 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     2041 atcacgaggc cctttcgtct tcaagaattc tcatgtttga cagcttatca tcgataagct
     2101 tcgaatggtg caaaaccttt cgcggtatgg catgatagcg cccggaagag agtcaattca
     2161 gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt
     2221 atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa
     2281 aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca caacaactgg
     2341 cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg cacgcgccgt
     2401 cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt
     2461 cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc
     2521 aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc attgctgtgg
     2581 aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag acacccatca
     2641 acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat ctggtcgcat
     2701 tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc
     2761 gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata gcggaacggg
     2821 aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg aatgagggca
     2881 tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca atgcgcgcca
     2941 ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac gacgataccg
     3001 aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg
     3061 ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc
     3121 agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg
     3181 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     3241 aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta ggcacaattc
     3301 tcatgtttga cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc
     3361 catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca
     3421 aggcgcactc ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat
     3481 attctgaaat gagctgctcg agttgacaat taatcatcgg ctcgtataat gtgtggaatt
     3541 gtgagcggat aacaatttca cacaggaaac agccagtccg tttaggtgtt ttcacgagcc
     3601 attcaccaac aaggagttaa ttaactatca tagccggcca ccatcaccat caccattaga
     3661 ctagtgagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag
     3721 aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg
     3781 atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta
     3841 ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca atgaaagacg
     3901 gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg
     3961 aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat
     4021 attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg
     4081 agaatatgtt tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg
     4141 tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat tatacgcaag
     4201 gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt gatggcttcc
     4261 atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag ggcggggcgt
     4321 aatgactagt gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
     4381 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
     4441 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     4501 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     4561 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     4621 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     4681 ctgacggatg gcctttttgc g
//
