LOCUS       Exported                6580 bp DNA     circular SYN 13-JUL-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6580)
  AUTHORS   Institut Fresnel
  TITLE     Direct Submission
  JOURNAL   Exported Jul 27, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6580
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     81..100
                     /label=T7
                     /note="color: black; sequence: TAATACGACTCACTATAGGG"
     promoter        81..99
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    100..124
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
                     /note="color: #31849b"
     primer_bind     117..177
                     /label=SpeISept9_i3FOR
                     /note="color: black; sequence: 
                     ACAATTCCCCACTAGTAATAATTTTGTTTAACTTTAAGAAGGAGATATACAtATGGAGA
                     GG"
     primer_bind     complement(127..142)
                     /label=SpeISept9TEVREV
                     /note="color: black; sequence: 
                     CAAAATTATTACTAGTTTATTTTTCGAACTGCGGGTGGCTCCAGCCGCTGCCGCTGCCC
                     TGGAAGTAAAGGTTTTCCATCTCTGGGGCTTCTGGC"
     RBS             139..161
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     primer_bind     155..171
                     /label=NdeISept7i1FOR
                     /note="color: black; sequence: 
                     AAGGAGATATACAtatgtcggtcagtgcgagatccgctgctgctgaggagaggagcgtc
                     aacagcagcaccATGGTAGCTCAACAGAAGAACCTTG"
     CDS             169..1929
                     /codon_start=1
                     /label=SEPT9_i3-TEV-Strep
                     /note="HuSep9"
                     /note="This feature has 11 segments:
                         1:  169 ..  189 / #ff99cc / SEPT9_i3-specific
                         2:  190 ..  606 / #ccffcc / SEPT9 NTE
                         3:  607 ..  609 / #ff00ff / onset of SEPT9_i4
                         4:  610 ..  867 / #ccffcc / SEPT9 NTE
                         5:  868 ..  870 / #ff00ff / onset of SEPT9_i5
                         6:  871 .. 1002 / #ccffcc / SEPT9 NTE
                         7: 1003 .. 1818 / #00ff00 / SEPT9 G
                         8: 1819 .. 1872 / #ccffcc / SEPT9 CTE
                         9: 1873 .. 1893 / #ff9900 / TEV site
                        10: 1894 .. 1905 / #00ff00
                        11: 1906 .. 1929 / #ff9900 / Strep tag"
                     /translation="MERDRISALKRSFEVEEVETPNSTPPRRVQTPLLRATVASSTQKF
                     QDLGVKNSEPSARHVDSLSQRSPKASLRRVELSGPKAAEPVSRRTELSIDISSKQVENA
                     GAIGPSRFGLKRAEVLGHKTPEPAPRRTEITIVKPQESAHRRMEPPASKVPEVPTAPAT
                     DAAPKRVEIQMPKPAEAPTAPSPAQTLENSEPAPVSQLQSRLEPKPQPPVAEATPRSQE
                     ATEAAPSCVGDMADTPRDAGLKQAPASRNEKAPVDFGYVGIDSILEQMRRKAMKQGFEF
                     NIMVVGQSGLGKSTLINTLFKSKISRKSVQPTSEERIPKTIEIKSITHDIEEKGVRMKL
                     TVIDTPGFGDHINNENCWQPIMKFINDQYEKYLQEEVNINRKKRIPDTRVHCCLYFIPA
                     TGHSLRPLDIEFMKRLSKVVNIVPVIAKADTLTLEERVHFKQRITADLLSNGIDVYPQK
                     EFDEDSEDRLVNEKFREMIPFAVVGSDHEYQVNGKRILGRKTKWGTIEVENTTHCEFAY
                     LRDLLIRTHMQNIKDITSSIHFEAYRVKRLNEGSSAMANGVEEKEPEAPEMENLYFQGS
                     GSGWSHPQFEK"
     primer_bind     355..372
                     /label=Sept9NFOR
                     /note="color: black; sequence: CTAAGCCAACGCTCCCCC"
     primer_bind     complement(815..832)
                     /label=Sept9NREV
                     /note="color: black; sequence: CCTGGCTCCGGGGTGTAG"
     primer_bind     1348..1367
                     /label=Sept9GFOR
                     /note="color: black; sequence: CTCTACTTCATCCCCGCCAC"
     primer_bind     1550..1568
                     /label=Sept9GseqFOR
                     /note="color: black; sequence: ATGAGGACTCGGAGGACCG"
     primer_bind     complement(1615..1633)
                     /label=Sept9GREV
                     /note="color: black; sequence: GGTACTCGTGGTCACTGCC"
     primer_bind     complement(1854..1948)
                     /label=SpeISept9TEVREV
                     /note="color: black; sequence: 
                     CAAAATTATTACTAGTTTATTTTTCGAACTGCGGGTGGCTCCAGCCGCTGCCGCTGCCC
                     TGGAAGTAAAGGTTTTCCATCTCTGGGGCTTCTGGC"
     RBS             1945..1967
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     primer_bind     1961..2056
                     /label=NdeISept7i1FOR
                     /note="color: black; sequence: 
                     AAGGAGATATACAtatgtcggtcagtgcgagatccgctgctgctgaggagaggagcgtc
                     aacagcagcaccATGGTAGCTCAACAGAAGAACCTTG"
     CDS             1975..3288
                     /codon_start=1
                     /label=SEPT7
                     /note="HSep7"
                     /note="color: #00ff00"
                     /translation="MSVSARSAAAEERSVNSSTMVAQQKNLEGYVGFANLPNQVYRKSV
                     KRGFEFTLMVVGESGLGKSTLINSLFLTDLYSPEYPGPSHRIKKTVQVEQSKVLIKEGG
                     VQLLLTIVDTPGFGDAVDNSNCWQPVIDYIDSKFEDYLNAESRVNRRQMPDNRVQCCLY
                     FIAPSGHGLKPLDIEFMKRLHEKVNIIPLIAKADTLTPEECQQFKKQIMKEIQEHKIKI
                     YEFPETDDEEENKLVKKIKDRLPLAVVGSNTIIEVNGKRVRGRQYPWGVAEVENGEHCD
                     FTILRNMLIRTHMQDLKDVTNNVHYENYRSRKLAAVTYNGVDNNKNKGQLTKSPLAQME
                     EERREHVAKMKKMEMEMEQVFEMKVKEKVQKLKDSEAELQRRHEQMKKNLEAQHKELEE
                     KRRQFEDEKANWEAQQRILEQQNSSRTLEKNKKKGKIF"
     primer_bind     complement(2250..2272)
                     /label=Sept7REV2
                     /note="color: black; sequence: TTAAAACTTTGGATTGTTCCACC"
     primer_bind     2787..2807
                     /label=Sept7FOR
                     /note="color: black; sequence: TGCTGAAGTTGAAAATGGTGA"
     primer_bind     complement(3051..3072)
                     /label=Sept7REV
                     /note="color: black; sequence: TTTGACCTTCATCTCAAACACC"
     primer_bind     complement(3255..3322)
                     /label=NheISept7REV
                     /note="color: black; sequence: 
                     GCAGCCTAGGGCTAGCTCTAGACTATTAGGATCcTTAAAAGATCTTCCCTTTCTTCTTG
                     TTCTTTTCC; added: 2021-07-13"
     primer_bind     complement(3327..3345)
                     /label=T7term
                     /note="color: black; sequence: CTAGTTATTGCTCAGCGGT"
     terminator      3342..3389
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     CDS             complement(3560..4351)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /note="color: #ccffcc"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4352..4443)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(4491..5229)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
                     /note="color: #ffff00"
     primer_bind     4837..4856
                     /label=dual79i3CSseqFOR
                     /note="color: black; sequence: TATCTCGGCTCGGTGTAGGT"
     primer_bind     4952..4970
                     /label=79i3CSseqFOR2
                     /note="color: black; sequence: agcacggtaaaacgccact"
     primer_bind     complement(5219..5242)
                     /label=dual79i3CSseqREV
                     /note="color: black; sequence: gttcagtagaaaagatcaaaggat"
     CDS             complement(5439..6521)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /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)."
                     /note="color: #993366"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(join(6522..6580,1..19))
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
       61 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
      121 ttccccacta gtaataattt tgtttaactt taagaaggag atatacatat ggagagggac
      181 cggatctcag ccttgaaaag atcttttgag gtcgaggagg tcgagacacc caactccacc
      241 ccaccccgga gggtccagac tcccctactc cgagccactg tggccagctc cacccagaaa
      301 ttccaggacc tgggcgtgaa gaactcagaa ccctcggccc gccatgtgga ctccctaagc
      361 caacgctccc ccaaggcgtc cctgcggagg gtggagctct cgggccccaa ggcggccgag
      421 ccggtgtccc ggcgcactga gctgtccatt gacatctcgt ccaagcaggt ggagaacgcc
      481 ggggccatcg gcccgtcccg gttcgggctc aagagggccg aggtgttggg ccacaagacg
      541 ccagaaccgg cccctcggag gacggagatc accatcgtca aaccccagga gtcagcccac
      601 cggaggatgg agccccctgc ctccaaggtc cccgaggtgc ccactgcccc tgccaccgac
      661 gcagccccca agagggtgga gatccagatg cccaagcctg ctgaggcgcc caccgccccc
      721 agcccagccc agaccttgga aaattcagag cctgcccctg tgtctcagct gcagagcagg
      781 ctggagccca agccccagcc ccctgtggct gaggctacac cccggagcca ggaggccact
      841 gaggcggctc ccagctgcgt tggcgacatg gccgacaccc ccagagatgc cgggctcaag
      901 caggcgcctg catcacggaa cgagaaggcc ccggtggact tcggctacgt ggggattgac
      961 tccatcctgg agcagatgcg ccggaaggcc atgaagcagg gcttcgagtt caacatcatg
     1021 gtggtcgggc agagcggctt gggtaaatcc accttaatca acaccctctt caaatccaaa
     1081 atcagccgga agtcggtgca gcccacctca gaggagcgca tccccaagac catcgagatc
     1141 aagtccatca cgcacgatat tgaggagaaa ggcgtccgga tgaagctgac agtgattgac
     1201 acaccagggt tcggggacca catcaacaac gagaactgct ggcagcccat catgaagttc
     1261 atcaatgacc agtacgagaa atacctgcag gaggaggtca acatcaaccg caagaagcgc
     1321 atcccggaca cccgcgtcca ctgctgcctc tacttcatcc ccgccaccgg ccactccctc
     1381 aggcccctgg acatcgagtt tatgaaacgc ctgagcaagg tggtcaacat cgtccctgtc
     1441 atcgccaagg cggacacact caccctggag gagagggtcc acttcaaaca gcggatcacc
     1501 gcagacctgc tgtccaacgg catcgacgtg tacccccaga aggaatttga tgaggactcg
     1561 gaggaccggc tggtgaacga gaagttccgg gagatgatcc catttgctgt ggtgggcagt
     1621 gaccacgagt accaggtcaa cggcaagagg atccttggga ggaagaccaa gtggggtacc
     1681 atcgaagttg aaaacaccac acactgtgag tttgcctacc tgcgggacct tctcatcagg
     1741 acgcacatgc agaacatcaa ggacatcacc agcagcatcc acttcgaggc gtaccgtgtg
     1801 aagcgcctca acgagggcag cagcgccatg gccaacggcg tggaggagaa ggagccagaa
     1861 gccccagaga tggaaaacct ttacttccag ggcagcggca gcggctggag ccacccgcag
     1921 ttcgaaaaat aaactagtaa taattttgtt taactttaag aaggagatat acatatgtcg
     1981 gtcagtgcga gatccgctgc tgctgaggag aggagcgtca acagcagcac catggtagct
     2041 caacagaaga accttgaagg ctatgtggga tttgccaatc tcccaaatca agtatacaga
     2101 aaatcggtga agagaggttt tgaattcacg cttatggtag tgggtgaatc tggattggga
     2161 aagtcgacat taatcaactc attattcctc acagatttgt attctccaga gtatccaggt
     2221 ccttctcata gaattaaaaa gactgtacag gtggaacaat ccaaagtttt aatcaaagaa
     2281 ggtggtgttc agttgctgct cacaatagtt gataccccag gatttggaga tgcagtggat
     2341 aatagtaatt gctggcagcc tgttatcgac tacattgata gtaaatttga ggactaccta
     2401 aatgcagaat cacgagtgaa cagacgtcag atgcctgata acagggtgca gtgttgttta
     2461 tacttcattg ctccttcagg acatggactt aaaccattgg atattgagtt tatgaagcgt
     2521 ttgcatgaaa aagtgaatat catcccactt attgccaaag cagacacact cacaccagag
     2581 gaatgccaac agtttaaaaa acagataatg aaagaaatcc aagaacataa aattaaaata
     2641 tacgaatttc cagaaacaga tgatgaagaa gaaaataaac ttgttaaaaa gataaaggac
     2701 cgtttacctc ttgctgtggt aggtagtaat actatcattg aagttaatgg caaaagggtc
     2761 agaggaaggc agtatccttg gggtgttgct gaagttgaaa atggtgaaca ttgtgatttt
     2821 acaatcctaa gaaatatgtt gataagaaca cacatgcagg acttgaaaga tgttactaat
     2881 aatgtccact atgagaacta cagaagcaga aaacttgcag ctgtgactta taatggagtt
     2941 gataacaaca agaataaagg gcagctgact aagagccctc tggcacaaat ggaagaagaa
     3001 agaagggagc atgtagctaa aatgaagaag atggagatgg agatggagca ggtgtttgag
     3061 atgaaggtca aagaaaaagt tcaaaaactg aaggactctg aagctgagct ccagcggcgc
     3121 catgagcaaa tgaaaaagaa tttggaagca cagcacaaag aattggagga aaaacgtcgt
     3181 cagttcgagg atgagaaagc aaactgggaa gctcaacaac gtattttaga acaacagaac
     3241 tcttcaagaa ccttggaaaa gaacaagaag aaagggaaga tcttttaagg atcctaatag
     3301 tctagagcta gccctaggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     3361 cctctaaacg ggtcttgagg ggttttttgc tgaaacctca ggcatttgag aagcacacgg
     3421 tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa gcggctattt
     3481 aacgaccctg ccctgaaccg acgaccgggt catcgtggcc ggatcttgcg gcccctcggc
     3541 ttgaacgaat tgttagacat tatttgccga ctaccttggt gatctcgcct ttcacgtagt
     3601 ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat
     3661 aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc
     3721 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg
     3781 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg
     3841 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct
     3901 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca
     3961 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt
     4021 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa
     4081 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca
     4141 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca
     4201 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta
     4261 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag
     4321 tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcaatat tattgaagca
     4381 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4441 aaatagctag gtcactcggt cgctacgctc cgggcgtgag actgcggcgg gcgctgcgga
     4501 cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg tgaggcaaaa
     4561 cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca gagttcacat
     4621 aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg aatctgacag
     4681 tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt cgctccctct
     4741 tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt tctcccttac
     4801 gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt gtaggtcgtt
     4861 cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg cgccttatcc
     4921 ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact ggcagcagcc
     4981 attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg ctcttgaagt
     5041 gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc tgcgaaagcc
     5101 agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc acctccccag
     5161 gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc tcaagaagat
     5221 cctttgatct tttctactga accgctcttg atttcagtgc aatttatctc ttcaaatgta
     5281 gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc atgccccgcg
     5341 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     5401 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5461 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5521 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     5581 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     5641 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     5701 tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc
     5761 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     5821 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     5881 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     5941 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     6001 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     6061 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     6121 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     6181 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     6241 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     6301 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     6361 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     6421 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     6481 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     6541 ctctcttccg ggcgctatca tgccataccg cgaaaggttt
//
