LOCUS       Exported                6634 bp DNA     circular SYN 13-JUL-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  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..6634
                     /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..180
                     /label=SpeISept9_i1FOR
                     /note="color: black; sequence: 
                     ACAATTCCCCACTAGTAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGAAGA
                     AGTCT"
     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..1983
                     /codon_start=1
                     /label=SEPT9_i1-TEV-Strep
                     /note="This feature has 11 segments:
                         1:  169 ..  243 / #ff99cc / SEPT9_i1-specific
                         2:  244 ..  660 / #ccffcc / SEPT9 NTE
                         3:  661 ..  663 / #ff00ff / onset of SEPT9_i4
                         4:  664 ..  921 / #ccffcc / SEPT9 NTE
                         5:  922 ..  924 / #ff00ff / onset of SEPT9_i5
                         6:  925 .. 1056 / #ccffcc / SEPT9 NTE
                         7: 1057 .. 1872 / #00ff00 / SEPT9 G
                         8: 1873 .. 1926 / #ccffcc / SEPT9 CTE
                         9: 1927 .. 1947 / #ff9900 / TEV site
                        10: 1948 .. 1959 / #00ff00
                        11: 1960 .. 1983 / #ff9900 / Strep tag"
                     /translation="MKKSYSGGTRTSSGRLRRLGDSSGPALKRSFEVEEVETPNSTPPR
                     RVQTPLLRATVASSTQKFQDLGVKNSEPSARHVDSLSQRSPKASLRRVELSGPKAAEPV
                     SRRTELSIDISSKQVENAGAIGPSRFGLKRAEVLGHKTPEPAPRRTEITIVKPQESAHR
                     RMEPPASKVPEVPTAPATDAAPKRVEIQMPKPAEAPTAPSPAQTLENSEPAPVSQLQSR
                     LEPKPQPPVAEATPRSQEATEAAPSCVGDMADTPRDAGLKQAPASRNEKAPVDFGYVGI
                     DSILEQMRRKAMKQGFEFNIMVVGQSGLGKSTLINTLFKSKISRKSVQPTSEERIPKTI
                     EIKSITHDIEEKGVRMKLTVIDTPGFGDHINNENCWQPIMKFINDQYEKYLQEEVNINR
                     KKRIPDTRVHCCLYFIPATGHSLRPLDIEFMKRLSKVVNIVPVIAKADTLTLEERVHFK
                     QRITADLLSNGIDVYPQKEFDEDSEDRLVNEKFREMIPFAVVGSDHEYQVNGKRILGRK
                     TKWGTIEVENTTHCEFAYLRDLLIRTHMQNIKDITSSIHFEAYRVKRLNEGSSAMANGV
                     EEKEPEAPEMENLYFQGSGSGWSHPQFEK"
     primer_bind     409..426
                     /label=Sept9NFOR
                     /note="color: black; sequence: CTAAGCCAACGCTCCCCC"
     primer_bind     complement(869..886)
                     /label=Sept9NREV
                     /note="color: black; sequence: CCTGGCTCCGGGGTGTAG"
     primer_bind     1402..1421
                     /label=Sept9GFOR
                     /note="color: black; sequence: CTCTACTTCATCCCCGCCAC"
     primer_bind     1604..1622
                     /label=Sept9GseqFOR
                     /note="color: black; sequence: ATGAGGACTCGGAGGACCG"
     primer_bind     complement(1669..1687)
                     /label=Sept9GREV
                     /note="color: black; sequence: GGTACTCGTGGTCACTGCC"
     primer_bind     complement(1908..2002)
                     /label=SpeISept9TEVREV
                     /note="color: black; sequence: 
                     CAAAATTATTACTAGTTTATTTTTCGAACTGCGGGTGGCTCCAGCCGCTGCCGCTGCCC
                     TGGAAGTAAAGGTTTTCCATCTCTGGGGCTTCTGGC"
     RBS             1999..2021
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     primer_bind     2015..2110
                     /label=NdeISept7i1FOR
                     /note="color: black; sequence: 
                     AAGGAGATATACAtatgtcggtcagtgcgagatccgctgctgctgaggagaggagcgtc
                     aacagcagcaccATGGTAGCTCAACAGAAGAACCTTG"
     CDS             2029..3342
                     /codon_start=1
                     /label=SEPT7
                     /note="HSep7"
                     /note="color: #00ff00"
                     /translation="MSVSARSAAAEERSVNSSTMVAQQKNLEGYVGFANLPNQVYRKSV
                     KRGFEFTLMVVGESGLGKSTLINSLFLTDLYSPEYPGPSHRIKKTVQVEQSKVLIKEGG
                     VQLLLTIVDTPGFGDAVDNSNCWQPVIDYIDSKFEDYLNAESRVNRRQMPDNRVQCCLY
                     FIAPSGHGLKPLDIEFMKRLHEKVNIIPLIAKADTLTPEECQQFKKQIMKEIQEHKIKI
                     YEFPETDDEEENKLVKKIKDRLPLAVVGSNTIIEVNGKRVRGRQYPWGVAEVENGEHCD
                     FTILRNMLIRTHMQDLKDVTNNVHYENYRSRKLAAVTYNGVDNNKNKGQLTKSPLAQME
                     EERREHVAKMKKMEMEMEQVFEMKVKEKVQKLKDSEAELQRRHEQMKKNLEAQHKELEE
                     KRRQFEDEKANWEAQQRILEQQNSSRTLEKNKKKGKIF"
     primer_bind     complement(2304..2326)
                     /label=Sept7REV2
                     /note="color: black; sequence: TTAAAACTTTGGATTGTTCCACC"
     primer_bind     2841..2861
                     /label=Sept7FOR
                     /note="color: black; sequence: TGCTGAAGTTGAAAATGGTGA"
     primer_bind     complement(3105..3126)
                     /label=Sept7REV
                     /note="color: black; sequence: TTTGACCTTCATCTCAAACACC"
     primer_bind     complement(3309..3376)
                     /label=NheISept7REV
                     /note="color: black; sequence: 
                     GCAGCCTAGGGCTAGCTCTAGACTATTAGGATCcTTAAAAGATCTTCCCTTTCTTCTTG
                     TTCTTTTCC; added: 2021-07-13"
     primer_bind     complement(3381..3399)
                     /label=T7term
                     /note="color: black; sequence: CTAGTTATTGCTCAGCGGT"
     terminator      3396..3443
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     CDS             complement(3614..4405)
                     /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(4406..4497)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(4545..5283)
                     /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     4891..4910
                     /label=dual79i3CSseqFOR
                     /note="color: black; sequence: TATCTCGGCTCGGTGTAGGT"
     primer_bind     5006..5024
                     /label=79i3CSseqFOR2
                     /note="color: black; sequence: agcacggtaaaacgccact"
     primer_bind     complement(5273..5296)
                     /label=dual79i3CSseqREV
                     /note="color: black; sequence: gttcagtagaaaagatcaaaggat"
     CDS             complement(5493..6575)
                     /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(6576..6634,1..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 gaagaagtct
      181 tactcaggag gcacgcggac ctccagtggc cggctccgga ggcttggtga ctccagtggc
      241 ccagccttaa aaagatcttt tgaggtcgag gaggtcgaga cacccaactc caccccaccc
      301 cggagggtcc agactcccct actccgagcc actgtggcca gctccaccca gaaattccag
      361 gacctgggcg tgaagaactc agaaccctcg gcccgccatg tggactccct aagccaacgc
      421 tcccccaagg cgtccctgcg gagggtggag ctctcgggcc ccaaggcggc cgagccggtg
      481 tcccggcgca ctgagctgtc cattgacatc tcgtccaagc aggtggagaa cgccggggcc
      541 atcggcccgt cccggttcgg gctcaagagg gccgaggtgt tgggccacaa gacgccagaa
      601 ccggcccctc ggaggacgga gatcaccatc gtcaaacccc aggagtcagc ccaccggagg
      661 atggagcccc ctgcctccaa ggtccccgag gtgcccactg cccctgccac cgacgcagcc
      721 cccaagaggg tggagatcca gatgcccaag cctgctgagg cgcccaccgc ccccagccca
      781 gcccagacct tggagaattc agagcctgcc cctgtgtctc agctgcagag caggctggag
      841 cccaagcccc agccccctgt ggctgaggct acaccccgga gccaggaggc cactgaggcg
      901 gctcccagct gcgttggcga catggccgac acccccagag atgccgggct caagcaggcg
      961 cctgcatcac ggaacgagaa ggccccggtg gacttcggct acgtggggat tgactccatc
     1021 ctggagcaga tgcgccggaa ggccatgaag cagggcttcg agttcaacat catggtggtc
     1081 gggcagagcg gcttgggtaa atccacctta atcaacaccc tcttcaaatc caaaatcagc
     1141 cggaagtcgg tgcagcccac ctcagaggag cgcatcccca agaccatcga gatcaagtcc
     1201 atcacgcacg atattgagga gaaaggcgtc cggatgaagc tgacagtgat tgacacacca
     1261 gggttcgggg accacatcaa caacgagaac tgctggcagc ccatcatgaa gttcatcaat
     1321 gaccagtacg agaaatacct gcaggaggag gtcaacatca accgcaagaa gcgcatcccg
     1381 gacacccgcg tccactgctg cctctacttc atccccgcca ccggccactc cctcaggccc
     1441 ctggacatcg agtttatgaa acgcctgagc aaggtggtca acatcgtccc tgtcatcgcc
     1501 aaggcggaca cactcaccct ggaggagagg gtccacttca aacagcggat caccgcagac
     1561 ctgctgtcca acggcatcga cgtgtacccc cagaaggaat ttgatgagga ctcggaggac
     1621 cggctggtga acgagaagtt ccgggagatg atcccatttg ctgtggtggg cagtgaccac
     1681 gagtaccagg tcaacggcaa gaggatcctt gggaggaaga ccaagtgggg taccatcgaa
     1741 gttgaaaaca ccacacactg tgagtttgcc tacctgcggg accttctcat caggacgcac
     1801 atgcagaaca tcaaggacat caccagcagc atccacttcg aggcgtaccg tgtgaagcgc
     1861 ctcaacgagg gcagcagcgc catggccaac ggcgtggagg agaaggagcc agaagcccca
     1921 gagatggaaa acctttactt ccagggcagc ggcagcggct ggagccaccc gcagttcgaa
     1981 aaataaacta gtaataattt tgtttaactt taagaaggag atatacatat gtcggtcagt
     2041 gcgagatccg ctgctgctga ggagaggagc gtcaacagca gcaccatggt agctcaacag
     2101 aagaaccttg aaggctatgt gggatttgcc aatctcccaa atcaagtata cagaaaatcg
     2161 gtgaagagag gttttgaatt cacgcttatg gtagtgggtg aatctggatt gggaaagtcg
     2221 acattaatca actcattatt cctcacagat ttgtattctc cagagtatcc aggtccttct
     2281 catagaatta aaaagactgt acaggtggaa caatccaaag ttttaatcaa agaaggtggt
     2341 gttcagttgc tgctcacaat agttgatacc ccaggatttg gagatgcagt ggataatagt
     2401 aattgctggc agcctgttat cgactacatt gatagtaaat ttgaggacta cctaaatgca
     2461 gaatcacgag tgaacagacg tcagatgcct gataacaggg tgcagtgttg tttatacttc
     2521 attgctcctt caggacatgg acttaaacca ttggatattg agtttatgaa gcgtttgcat
     2581 gaaaaagtga atatcatccc acttattgcc aaagcagaca cactcacacc agaggaatgc
     2641 caacagttta aaaaacagat aatgaaagaa atccaagaac ataaaattaa aatatacgaa
     2701 tttccagaaa cagatgatga agaagaaaat aaacttgtta aaaagataaa ggaccgttta
     2761 cctcttgctg tggtaggtag taatactatc attgaagtta atggcaaaag ggtcagagga
     2821 aggcagtatc cttggggtgt tgctgaagtt gaaaatggtg aacattgtga ttttacaatc
     2881 ctaagaaata tgttgataag aacacacatg caggacttga aagatgttac taataatgtc
     2941 cactatgaga actacagaag cagaaaactt gcagctgtga cttataatgg agttgataac
     3001 aacaagaata aagggcagct gactaagagc cctctggcac aaatggaaga agaaagaagg
     3061 gagcatgtag ctaaaatgaa gaagatggag atggagatgg agcaggtgtt tgagatgaag
     3121 gtcaaagaaa aagttcaaaa actgaaggac tctgaagctg agctccagcg gcgccatgag
     3181 caaatgaaaa agaatttgga agcacagcac aaagaattgg aggaaaaacg tcgtcagttc
     3241 gaggatgaga aagcaaactg ggaagctcaa caacgtattt tagaacaaca gaactcttca
     3301 agaaccttgg aaaagaacaa gaagaaaggg aagatctttt aaggatccta atagtctaga
     3361 gctagcccta ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     3421 aacgggtctt gaggggtttt ttgctgaaac ctcaggcatt tgagaagcac acggtcacac
     3481 tgcttccggt agtcaataaa ccggtaaacc agcaatagac ataagcggct atttaacgac
     3541 cctgccctga accgacgacc gggtcatcgt ggccggatct tgcggcccct cggcttgaac
     3601 gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa
     3661 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
     3721 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
     3781 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
     3841 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
     3901 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
     3961 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
     4021 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
     4081 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
     4141 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt
     4201 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
     4261 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
     4321 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     4381 cttcggcgat caccgcttcc ctcatactct tcctttttca atattattga agcatttatc
     4441 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4501 ctaggtcact cggtcgctac gctccgggcg tgagactgcg gcgggcgctg cggacacata
     4561 caaagttacc cacagattcc gtggataagc aggggactaa catgtgaggc aaaacagcag
     4621 ggccgcgccg gtggcgtttt tccataggct ccgccctcct gccagagttc acataaacag
     4681 acgcttttcc ggtgcatctg tgggagccgt gaggctcaac catgaatctg acagtacggg
     4741 cgaaacccga caggacttaa agatccccac cgtttccggc gggtcgctcc ctcttgcgct
     4801 ctcctgttcc gaccctgccg tttaccggat acctgttccg cctttctccc ttacgggaag
     4861 tgtggcgctt tctcatagct cacacactgg tatctcggct cggtgtaggt cgttcgctcc
     4921 aagctgggct gtaagcaaga actccccgtt cagcccgact gctgcgcctt atccggtaac
     4981 tgttcacttg agtccaaccc ggaaaagcac ggtaaaacgc cactggcagc agccattggt
     5041 aactgggagt tcgcagagga tttgtttagc taaacacgcg gttgctcttg aagtgtgcgc
     5101 caaagtccgg ctacactgga aggacagatt tggttgctgt gctctgcgaa agccagttac
     5161 cacggttaag cagttcccca actgacttaa ccttcgatca aaccacctcc ccaggtggtt
     5221 ttttcgttta cagggcaaaa gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5281 atcttttcta ctgaaccgct cttgatttca gtgcaattta tctcttcaaa tgtagcacct
     5341 gaagtcagcc ccatacgata taagttgtaa ttctcatgtt agtcatgccc cgcgcccacc
     5401 ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat
     5461 gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5521 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5581 ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc
     5641 gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa
     5701 tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat
     5761 cccactaccg agatgtccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg
     5821 cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc
     5881 atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc
     5941 ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag
     6001 acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc
     6061 tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg
     6121 tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca
     6181 tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg
     6241 tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg
     6301 gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg
     6361 gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc
     6421 acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc
     6481 gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca
     6541 tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct
     6601 tccgggcgct atcatgccat accgcgaaag gttt
//
