LOCUS       Exported                6169 bp DNA     circular SYN 27-JUL-2021
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6169)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 27, 2021 from SnapGene Viewer 5.3.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6169
                     /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..188
                     /label=SpeISept6FOR
                     /note="color: black; sequence: 
                     ACAATTCCCCACTAGTAATAATTTTGTTTAACTTTAAGAAGGAGATATACATATGGCAG
                     CGACCGATATAGC"
     primer_bind     complement(127..142)
                     /label=SpeISept6REV
                     /note="color: black; sequence: 
                     CAAAATTATTACTAGTCTATTAGGATCCTTAATTTTTCTTCTCTTTGTCTCTCTTCAGA
                     GTCTGTGAG"
     misc_feature    128..132
                     /label=SpeI
                     /note="color: #a6acb3"
     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..1452
                     /codon_start=1
                     /label=SEPT6
                     /note="HuSep6"
                     /note="color: #00ff00"
                     /translation="MAATDIARQVGEGCRTVPLAGHVGFDSLPDQLVNKSVSQGFCFNI
                     LCVGETGLGKSTLMDTLFNTKFEGEPATHTQPGVQLQSNTYDLQESNVRLKLTIVSTVG
                     FGDQINKEDSYKPIVEFIDAQFEAYLQEELKIRRVLHTYHDSRIHVCLYFIAPTGHSLK
                     SLDLVTMKKLDSKVNIIPIIAKADAISKSELTKFKIKITSELVSNGVQIYQFPTDDESV
                     AEINGTMNAHLPFAVIGSTEELKIGNKMMRARQYPWGTVQVENEAHCDFVKLREMLIRV
                     NMEDLREQTHTRHYELYRRCKLEEMGFKDTDPDSKPFSLQETYEAKRNEFLGELQKKEE
                     EMRQMFVQRVKEKEAELKEAEKELHEKFDRLKKLHQDEKKKLEDKKKSLDDEVNAFKQR
                     KTAAELLQSQGSQAGGSQTLKRDKEKKN"
     primer_bind     complement(788..808)
                     /label=Sept6REV2
                     /note="color: black; sequence: GATAGATCTGGACTCCGTTGC"
     primer_bind     928..945
                     /label=Sept6FOR2
                     /note="color: black; sequence: CGGCAGTATCCTTGGGGC"
     primer_bind     1128..1148
                     /label=SEP6FOR
                     /note="color: black; sequence: ACAGGAGACATATGAGGCCAA"
     primer_bind     complement(1193..1213)
                     /label=Sept6REV
                     /note="color: black; sequence: GCTGGACGAACATCTGTCTCA"
     primer_bind     complement(1413..1480)
                     /label=SpeISept6REV
                     /note="color: black; sequence: 
                     CAAAATTATTACTAGTCTATTAGGATCCTTAATTTTTCTTCTCTTTGTCTCTCTTCAGA
                     GTCTGTGAG"
     misc_feature    1465
                     /label=SpeI
                     /note="color: #a6acb3"
     RBS             1477..1499
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
                     /note="color: #a6acb3"
     primer_bind     1493..1588
                     /label=NdeISept7i1FOR
                     /note="color: black; sequence: 
                     AAGGAGATATACAtatgtcggtcagtgcgagatccgctgctgctgaggagaggagcgtc
                     aacagcagcaccATGGTAGCTCAACAGAAGAACCTTG"
     CDS             1507..2877
                     /codon_start=1
                     /label=SEPT7-TEV-Strep
                     /note="HSep7"
                     /note="This feature has 5 segments:
                        1: 1507 .. 2817 / #00ff00
                        2: 2818 .. 2838 / #ff9900 / TEV site
                        3: 2839 .. 2850 / #00ff00
                        4: 2851 .. 2874 / #ff9900 / Strep tag
                        5: 2875 .. 2877 / #00ff00"
                     /translation="MSVSARSAAAEERSVNSSTMVAQQKNLEGYVGFANLPNQVYRKSV
                     KRGFEFTLMVVGESGLGKSTLINSLFLTDLYSPEYPGPSHRIKKTVQVEQSKVLIKEGG
                     VQLLLTIVDTPGFGDAVDNSNCWQPVIDYIDSKFEDYLNAESRVNRRQMPDNRVQCCLY
                     FIAPSGHGLKPLDIEFMKRLHEKVNIIPLIAKADTLTPEECQQFKKQIMKEIQEHKIKI
                     YEFPETDDEEENKLVKKIKDRLPLAVVGSNTIIEVNGKRVRGRQYPWGVAEVENGEHCD
                     FTILRNMLIRTHMQDLKDVTNNVHYENYRSRKLAAVTYNGVDNNKNKGQLTKSPLAQME
                     EERREHVAKMKKMEMEMEQVFEMKVKEKVQKLKDSEAELQRRHEQMKKNLEAQHKELEE
                     KRRQFEDEKANWEAQQRILEQQNSSRTLEKNKKKGKIFENLYFQGSGSGWSHPQFEK"
     primer_bind     complement(1782..1804)
                     /label=Sept7REV2
                     /note="color: black; sequence: TTAAAACTTTGGATTGTTCCACC"
     primer_bind     2319..2339
                     /label=Sept7FOR
                     /note="color: black; sequence: TGCTGAAGTTGAAAATGGTGA"
     primer_bind     complement(2583..2604)
                     /label=Sept7REV
                     /note="color: black; sequence: TTTGACCTTCATCTCAAACACC"
     primer_bind     complement(2787..2911)
                     /label=NheISept7TEVREV
                     /note="color: black; sequence: 
                     GCAGCCTAGGGCTAGCTCTAGACTATTAGGATCcTTATTTTTCGAACTGCGGGTGGCTC
                     CAGCCGCTGCCGCTGCCCTGGAAGTAAAGGTTTTCAAAGATCTTCCCTTTCTTCTTGTT
                     CTTTTCC"
     primer_bind     complement(2916..2934)
                     /label=T7term
                     /note="color: black; sequence: CTAGTTATTGCTCAGCGGT"
     terminator      2931..2978
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     CDS             complement(3149..3940)
                     /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(3941..4032)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(4080..4818)
                     /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     4426..4445
                     /label=dual79i3CSseqFOR
                     /note="color: black; sequence: TATCTCGGCTCGGTGTAGGT"
     primer_bind     4541..4559
                     /label=79i3CSseqFOR2
                     /note="color: black; sequence: agcacggtaaaacgccact"
     primer_bind     complement(4808..4831)
                     /label=dual79i3CSseqREV
                     /note="color: black; sequence: gttcagtagaaaagatcaaaggat"
     CDS             complement(5028..6110)
                     /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(6111..6169,1..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 ggcagcgacc
      181 gatatagctc gccaggtggg tgaaggttgc cgaactgtcc ccctggctgg acatgtgggg
      241 tttgacagct tgcctgacca gctggtgaat aagtccgtca gccagggctt ctgcttcaac
      301 atcctgtgcg tgggagagac aggtttgggc aagtccaccc tcatggacac cctgttcaac
      361 accaaattcg aaggggagcc agccacccac acacagccgg gtgtccagct ccagtctaat
      421 acctatgacc tccaagagag caacgtgagg ctaaagctca cgatcgttag cacagttggc
      481 tttggggacc agatcaacaa agaggacagc tacaagccta tcgtggaatt catcgatgca
      541 caattcgagg cctacctgca ggaagagcta aagatccgaa gagtgctaca cacctaccat
      601 gactcccgaa tccatgtctg cttgtatttc attgccccca cgggtcattc cctgaagtct
      661 ctggacctag tgactatgaa gaagctggac agtaaggtga acatcatccc catcattgcc
      721 aaagcagatg ccatttcgaa gagtgagcta acaaagttca aaatcaaaat caccagcgag
      781 cttgtcagca acggagtcca gatctatcag tttcctacag atgatgagtc ggtggcagag
      841 atcaatggaa ccatgaacgc ccacctgccg tttgctgtca ttggcagcac agaagaactg
      901 aagataggca acaagatgat gagggcgcgg cagtatcctt ggggcactgt gcaggttgaa
      961 aacgaggccc actgcgactt tgtgaagctg cgggagatgc tgattcgggt caacatggag
     1021 gatctgcggg agcagaccca cacccggcac tatgagctgt atcgccgctg taagctggag
     1081 gagatgggct tcaaggacac cgaccctgac agcaaaccct tcagtttaca ggagacatat
     1141 gaggccaaaa ggaacgagtt cctaggggaa ctccagaaaa aagaagagga gatgagacag
     1201 atgttcgtcc agcgagtcaa agagaaagaa gcggagctca aagaggcaga gaaagagctg
     1261 cacgagaagt ttgaccgtct gaagaaactg caccaggacg agaagaagaa actggaggat
     1321 aagaagaaat ccctggatga tgaagtgaat gctttcaagc aaagaaagac ggcggctgag
     1381 ctgctccagt cccagggctc ccaggctgga ggctcacaga ctctgaagag agacaaagag
     1441 aagaaaaatt aaggatccta atagactagt aataattttg tttaacttta agaaggagat
     1501 atacatatgt cggtcagtgc gagatccgct gctgctgagg agaggagcgt caacagcagc
     1561 accatggtag ctcaacagaa gaaccttgaa ggctatgtgg gatttgccaa tctcccaaat
     1621 caagtataca gaaaatcggt gaagagaggt tttgaattca cgcttatggt agtgggtgaa
     1681 tctggattgg gaaagtcgac attaatcaac tcattattcc tcacagattt gtattctcca
     1741 gagtatccag gtccttctca tagaattaaa aagactgtac aggtggaaca atccaaagtt
     1801 ttaatcaaag aaggtggtgt tcagttgctg ctcacaatag ttgatacccc aggatttgga
     1861 gatgcagtgg ataatagtaa ttgctggcag cctgttatcg actacattga tagtaaattt
     1921 gaggactacc taaatgcaga atcacgagtg aacagacgtc agatgcctga taacagggtg
     1981 cagtgttgtt tatacttcat tgctccttca ggacatggac ttaaaccatt ggatattgag
     2041 tttatgaagc gtttgcatga aaaagtgaat atcatcccac ttattgccaa agcagacaca
     2101 ctcacaccag aggaatgcca acagtttaaa aaacagataa tgaaagaaat ccaagaacat
     2161 aaaattaaaa tatacgaatt tccagaaaca gatgatgaag aagaaaataa acttgttaaa
     2221 aagataaagg accgtttacc tcttgctgtg gtaggtagta atactatcat tgaagttaat
     2281 ggcaaaaggg tcagaggaag gcagtatcct tggggtgttg ctgaagttga aaatggtgaa
     2341 cattgtgatt ttacaatcct aagaaatatg ttgataagaa cacacatgca ggacttgaaa
     2401 gatgttacta ataatgtcca ctatgagaac tacagaagca gaaaacttgc agctgtgact
     2461 tataatggag ttgataacaa caagaataaa gggcagctga ctaagagccc tctggcacaa
     2521 atggaagaag aaagaaggga gcatgtagct aaaatgaaga agatggagat ggagatggag
     2581 caggtgtttg agatgaaggt caaagaaaaa gttcaaaaac tgaaggactc tgaagctgag
     2641 ctccagcggc gccatgagca aatgaaaaag aatttggaag cacagcacaa agaattggag
     2701 gaaaaacgtc gtcagttcga ggatgagaaa gcaaactggg aagctcaaca acgtatttta
     2761 gaacaacaga actcttcaag aaccttggaa aagaacaaga agaaagggaa gatctttgaa
     2821 aacctttact tccagggcag cggcagcggc tggagccacc cgcagttcga aaaataagga
     2881 tcctaatagt ctagagctag ccctaggctg ctgccaccgc tgagcaataa ctagcataac
     2941 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag gcatttgaga
     3001 agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa tagacataag
     3061 cggctattta acgaccctgc cctgaaccga cgaccgggtc atcgtggccg gatcttgcgg
     3121 cccctcggct tgaacgaatt gttagacatt atttgccgac taccttggtg atctcgcctt
     3181 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
     3241 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
     3301 ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc
     3361 aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt
     3421 tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa
     3481 agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca
     3541 agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc
     3601 gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt
     3661 cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg
     3721 aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca
     3781 ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt
     3841 acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg
     3901 atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt
     3961 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4021 aaaataaaca aatagctagg tcactcggtc gctacgctcc gggcgtgaga ctgcggcggg
     4081 cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg actaacatgt
     4141 gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc ctcctgccag
     4201 agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc tcaaccatga
     4261 atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt ccggcgggtc
     4321 gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg ttccgccttt
     4381 ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct cggctcggtg
     4441 taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc cgactgctgc
     4501 gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa aacgccactg
     4561 gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac acgcggttgc
     4621 tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt gctgtgctct
     4681 gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc gatcaaacca
     4741 cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa aaaaggatct
     4801 caagaagatc ctttgatctt ttctactgaa ccgctcttga tttcagtgca atttatctct
     4861 tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc atgttagtca
     4921 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     4981 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     5041 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5101 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     5161 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     5221 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     5281 tcggtatcgt cgtatcccac taccgagatg tccgcaccaa cgcgcagccc ggactcggta
     5341 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     5401 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     5461 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     5521 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     5581 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     5641 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     5701 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     5761 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     5821 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     5881 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     5941 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     6001 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     6061 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     6121 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggttt
//
