LOCUS       Exported                5881 bp DNA     circular SYN 13-JUL-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5881)
  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..5881
                     /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..185
                     /label=SpeISept9i5FOR
                     /note="color: black; sequence: 
                     ACAATTCCCCACTAGTAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGGCCG
                     ACACCCCCAG"
     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..1173
                     /codon_start=1
                     /label=SEPT9_i5-TEV-Strep
                     /note="This feature has 3 segments:
                        1:  169 ..  303 / #ccffcc / SEPT9 NTE
                        2:  304 .. 1119 / #00ff00 / SEPT9 G
                        3: 1120 .. 1173 / #ccffcc / SEPT9 CTE"
                     /translation="MADTPRDAGLKQAPASRNEKAPVDFGYVGIDSILEQMRRKAMKQG
                     FEFNIMVVGQSGLGKSTLINTLFKSKISRKSVQPTSEERIPKTIEIKSITHDIEEKGVR
                     MKLTVIDTPGFGDHINNENCWQPIMKFINDQYEKYLQEEVNINRKKRIPDTRVHCCLYF
                     IPATGHSLRPLDIEFMKRLSKVVNIVPVIAKADTLTLEERVHFKQRITADLLSNGIDVY
                     PQKEFDEDSEDRLVNEKFREMIPFAVVGSDHEYQVNGKRILGRKTKWGTIEVENTTHCE
                     FAYLRDLLIRTHMQNIKDITSSIHFEAYRVKRLNEGSSAMANGVEEKEPEAPEM"
     primer_bind     649..668
                     /label=Sept9GFOR
                     /note="color: black; sequence: CTCTACTTCATCCCCGCCAC"
     primer_bind     851..869
                     /label=Sept9GseqFOR
                     /note="color: black; sequence: ATGAGGACTCGGAGGACCG"
     primer_bind     complement(916..934)
                     /label=Sept9GREV
                     /note="color: black; sequence: GGTACTCGTGGTCACTGCC"
     primer_bind     complement(1155..1249)
                     /label=SpeISept9TEVREV
                     /note="color: black; sequence: 
                     CAAAATTATTACTAGTTTATTTTTCGAACTGCGGGTGGCTCCAGCCGCTGCCGCTGCCC
                     TGGAAGTAAAGGTTTTCCATCTCTGGGGCTTCTGGC"
     CDS             1174..1230
                     /codon_start=1
                     /label=Sept9_i1-Strep
                     /note="This feature has 3 segments:
                        1: 1174 .. 1194 / #ff9900 / TEV site
                        2: 1195 .. 1206 / #00ff00
                        3: 1207 .. 1230 / #ff9900 / Strep tag"
                     /translation="ENLYFQGSGSGWSHPQFEK"
     RBS             1246..1268
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     primer_bind     1262..1357
                     /label=NdeISept7i1FOR
                     /note="color: black; sequence: 
                     AAGGAGATATACAtatgtcggtcagtgcgagatccgctgctgctgaggagaggagcgtc
                     aacagcagcaccATGGTAGCTCAACAGAAGAACCTTG"
     CDS             1276..2589
                     /codon_start=1
                     /label=SEPT7
                     /note="HSep7"
                     /note="color: #00ff00"
                     /translation="MSVSARSAAAEERSVNSSTMVAQQKNLEGYVGFANLPNQVYRKSV
                     KRGFEFTLMVVGESGLGKSTLINSLFLTDLYSPEYPGPSHRIKKTVQVEQSKVLIKEGG
                     VQLLLTIVDTPGFGDAVDNSNCWQPVIDYIDSKFEDYLNAESRVNRRQMPDNRVQCCLY
                     FIAPSGHGLKPLDIEFMKRLHEKVNIIPLIAKADTLTPEECQQFKKQIMKEIQEHKIKI
                     YEFPETDDEEENKLVKKIKDRLPLAVVGSNTIIEVNGKRVRGRQYPWGVAEVENGEHCD
                     FTILRNMLIRTHMQDLKDVTNNVHYENYRSRKLAAVTYNGVDNNKNKGQLTKSPLAQME
                     EERREHVAKMKKMEMEMEQVFEMKVKEKVQKLKDSEAELQRRHEQMKKNLEAQHKELEE
                     KRRQFEDEKANWEAQQRILEQQNSSRTLEKNKKKGKIF"
     primer_bind     complement(1551..1573)
                     /label=Sept7REV2
                     /note="color: black; sequence: TTAAAACTTTGGATTGTTCCACC"
     primer_bind     2088..2108
                     /label=Sept7FOR
                     /note="color: black; sequence: TGCTGAAGTTGAAAATGGTGA"
     primer_bind     complement(2352..2373)
                     /label=Sept7REV
                     /note="color: black; sequence: TTTGACCTTCATCTCAAACACC"
     primer_bind     complement(2556..2623)
                     /label=NheISept7REV
                     /note="color: black; sequence: 
                     GCAGCCTAGGGCTAGCTCTAGACTATTAGGATCcTTAAAAGATCTTCCCTTTCTTCTTG
                     TTCTTTTCC; added: 2021-07-13"
     primer_bind     complement(2628..2646)
                     /label=T7term
                     /note="color: black; sequence: CTAGTTATTGCTCAGCGGT"
     terminator      2643..2690
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     CDS             complement(2861..3652)
                     /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(3653..3744)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(3792..4530)
                     /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     4138..4157
                     /label=dual79i3CSseqFOR
                     /note="color: black; sequence: TATCTCGGCTCGGTGTAGGT"
     primer_bind     4253..4271
                     /label=79i3CSseqFOR2
                     /note="color: black; sequence: agcacggtaaaacgccact"
     primer_bind     complement(4520..4543)
                     /label=dual79i3CSseqREV
                     /note="color: black; sequence: gttcagtagaaaagatcaaaggat"
     CDS             complement(4740..5822)
                     /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"
     primer_bind     complement(5306..5316)
                     /label=Sept9sfGFPFOR
                     /note="color: black; sequence: AGAGATGGTGTCCAAGGGCGAGGAGC"
     promoter        complement(join(5823..5881,1..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 ggccgacacc
      181 cccagagatg ccgggctcaa gcaggcgcct gcatcacgga acgagaaggc cccggtggac
      241 ttcggctacg tggggattga ctccatcctg gagcagatgc gccggaaggc catgaagcag
      301 ggcttcgagt tcaacatcat ggtggtcggg cagagcggct tgggtaaatc caccttaatc
      361 aacaccctct tcaaatccaa aatcagccgg aagtcggtgc agcccacctc agaggagcgc
      421 atccccaaga ccatcgagat caagtccatc acgcacgata ttgaggagaa aggcgtccgg
      481 atgaagctga cagtgattga cacaccaggg ttcggggacc acatcaacaa cgagaactgc
      541 tggcagccca tcatgaagtt catcaatgac cagtacgaga aatacctgca ggaggaggtc
      601 aacatcaacc gcaagaagcg catcccggac acccgcgtcc actgctgcct ctacttcatc
      661 cccgccaccg gccactccct caggcccctg gacatcgagt ttatgaaacg cctgagcaag
      721 gtggtcaaca tcgtccctgt catcgccaag gcggacacac tcaccctgga ggagagggtc
      781 cacttcaaac agcggatcac cgcagacctg ctgtccaacg gcatcgacgt gtacccccag
      841 aaggaatttg atgaggactc ggaggaccgg ctggtgaacg agaagttccg ggagatgatc
      901 ccatttgctg tggtgggcag tgaccacgag taccaggtca acggcaagag gatccttggg
      961 aggaagacca agtggggtac catcgaagtt gaaaacacca cacactgtga gtttgcctac
     1021 ctgcgggacc ttctcatcag gacgcacatg cagaacatca aggacatcac cagcagcatc
     1081 cacttcgagg cgtaccgtgt gaagcgcctc aacgagggca gcagcgccat ggccaacggc
     1141 gtggaggaga aggagccaga agccccagag atggaaaacc tttacttcca gggcagcggc
     1201 agcggctgga gccacccgca gttcgaaaaa taaactagta ataattttgt ttaactttaa
     1261 gaaggagata tacatatgtc ggtcagtgcg agatccgctg ctgctgagga gaggagcgtc
     1321 aacagcagca ccatggtagc tcaacagaag aaccttgaag gctatgtggg atttgccaat
     1381 ctcccaaatc aagtatacag aaaatcggtg aagagaggtt ttgaattcac gcttatggta
     1441 gtgggtgaat ctggattggg aaagtcgaca ttaatcaact cattattcct cacagatttg
     1501 tattctccag agtatccagg tccttctcat agaattaaaa agactgtaca ggtggaacaa
     1561 tccaaagttt taatcaaaga aggtggtgtt cagttgctgc tcacaatagt tgatacccca
     1621 ggatttggag atgcagtgga taatagtaat tgctggcagc ctgttatcga ctacattgat
     1681 agtaaatttg aggactacct aaatgcagaa tcacgagtga acagacgtca gatgcctgat
     1741 aacagggtgc agtgttgttt atacttcatt gctccttcag gacatggact taaaccattg
     1801 gatattgagt ttatgaagcg tttgcatgaa aaagtgaata tcatcccact tattgccaaa
     1861 gcagacacac tcacaccaga ggaatgccaa cagtttaaaa aacagataat gaaagaaatc
     1921 caagaacata aaattaaaat atacgaattt ccagaaacag atgatgaaga agaaaataaa
     1981 cttgttaaaa agataaagga ccgtttacct cttgctgtgg taggtagtaa tactatcatt
     2041 gaagttaatg gcaaaagggt cagaggaagg cagtatcctt ggggtgttgc tgaagttgaa
     2101 aatggtgaac attgtgattt tacaatccta agaaatatgt tgataagaac acacatgcag
     2161 gacttgaaag atgttactaa taatgtccac tatgagaact acagaagcag aaaacttgca
     2221 gctgtgactt ataatggagt tgataacaac aagaataaag ggcagctgac taagagccct
     2281 ctggcacaaa tggaagaaga aagaagggag catgtagcta aaatgaagaa gatggagatg
     2341 gagatggagc aggtgtttga gatgaaggtc aaagaaaaag ttcaaaaact gaaggactct
     2401 gaagctgagc tccagcggcg ccatgagcaa atgaaaaaga atttggaagc acagcacaaa
     2461 gaattggagg aaaaacgtcg tcagttcgag gatgagaaag caaactggga agctcaacaa
     2521 cgtattttag aacaacagaa ctcttcaaga accttggaaa agaacaagaa gaaagggaag
     2581 atcttttaag gatcctaata gtctagagct agccctaggc tgctgccacc gctgagcaat
     2641 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaacctc
     2701 aggcatttga gaagcacacg gtcacactgc ttccggtagt caataaaccg gtaaaccagc
     2761 aatagacata agcggctatt taacgaccct gccctgaacc gacgaccggg tcatcgtggc
     2821 cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca ttatttgccg actaccttgg
     2881 tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc gaggccaagc
     2941 gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc tgatactggg
     3001 ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt ttgccggtta
     3061 ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg ccagcccagt
     3121 cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga tcctgttcag
     3181 gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta tgttctcttg
     3241 cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag atacctgcaa
     3301 gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga taacgccacg
     3361 gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc tcgctctctc
     3421 caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt gtttcatcaa
     3481 gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg ccgccatcca
     3541 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc
     3601 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atactcttcc
     3661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3721 aatgtattta gaaaaataaa caaatagcta ggtcactcgg tcgctacgct ccgggcgtga
     3781 gactgcggcg ggcgctgcgg acacatacaa agttacccac agattccgtg gataagcagg
     3841 ggactaacat gtgaggcaaa acagcagggc cgcgccggtg gcgtttttcc ataggctccg
     3901 ccctcctgcc agagttcaca taaacagacg cttttccggt gcatctgtgg gagccgtgag
     3961 gctcaaccat gaatctgaca gtacgggcga aacccgacag gacttaaaga tccccaccgt
     4021 ttccggcggg tcgctccctc ttgcgctctc ctgttccgac cctgccgttt accggatacc
     4081 tgttccgcct ttctccctta cgggaagtgt ggcgctttct catagctcac acactggtat
     4141 ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta agcaagaact ccccgttcag
     4201 cccgactgct gcgccttatc cggtaactgt tcacttgagt ccaacccgga aaagcacggt
     4261 aaaacgccac tggcagcagc cattggtaac tgggagttcg cagaggattt gtttagctaa
     4321 acacgcggtt gctcttgaag tgtgcgccaa agtccggcta cactggaagg acagatttgg
     4381 ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag ttccccaact gacttaacct
     4441 tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat tacgcgcaga
     4501 aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctctt gatttcagtg
     4561 caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa gttgtaattc
     4621 tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg
     4681 gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     4741 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4801 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     4861 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     4921 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa
     4981 catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc aacgcgcagc
     5041 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     5101 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     5161 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     5221 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     5281 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     5341 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     5401 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     5461 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     5521 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     5581 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     5641 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     5701 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     5761 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     5821 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     5881 t
//
