LOCUS       Exported                6985 bp DNA     circular SYN 14-MAY-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6985)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 13, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..20
                     /label=DuetUP1
                     /note="color: black; sequence: GGATCTCGACGCTCTCCCT"
     promoter        45..63
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    64..88
                     /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"
     RBS             119..124
                     /note="color: #c0c0c0"
     CDS             132..134
                     /codon_start=1
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     primer_bind     154..197
                     /label=TIR1 F
                     /note="color: black; sequence: 
                     catcaccaagccaggGatccATGCAGAAGCGAATAGCCTTGTCG"
     CDS             174..1955
                     /codon_start=1
                     /label=AtTIR1
                     /note="color: #00ffff"
                     /translation="MQKRIALSFPEEVLEHVFSFIQLDKDRNSVSLVCKSWYEIERWCR
                     RKVFIGNCYAVSPATVIRRFPKVRSVELKGKPHFADFNLVPDGWGGYVYPWIEAMSSSY
                     TWLEEIRLKRMVVTDDCLELIAKSFKNFKVLVLSSCEGFSTDGLAAIAATCRNLKELDL
                     RESDVDDVSGHWLSHFPDTYTSLVSLNISCLASEVSFSALERLVTRCPNLKSLKLNRAV
                     PLEKLATLLQRAPQLEELGTGGYTAEVRPDVYSGLSVALSGCKELRCLSGFWDAVPAYL
                     PAVYSVCSRLTTLNLSYATVQSYDLVKLLCQCPKLQRLWVLDYIEDAGLEVLASTCKDL
                     RELRVFPSEPFVMEPNVALTEQGLVSVSMGCPKLESVLYFCRQMTNAALITIARNRPNM
                     TRFRLCIIEPKAPDYLTLEPLDIGFGAIVEHCKDLRRLSLSGLLTDKVFEYIGTYAKKM
                     EMLSVAFAGDSDLGMHHVLSGCDSLRKLEIRDCPFGDKALLANASKLETMRSLWMSSCS
                     VSFGACKLLGQKMPKLNVEVIDERGAPDSRPESCPVERVFIYRTVAGPRFDMPGFVWNM
                     DQDSTMRFSRQIITTNGL"
     primer_bind     complement(1927..1971)
                     /label=TIR1 R
                     /note="color: black; sequence: 
                     TATCGAGCTCGGATCCTAATCCGTTAGTAGTAATGATTTGCCTGG"
     CDS             1983..2042
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=2¡ÁMyc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDLEQKLISEEDL"
     primer_bind     complement(2105..2124)
                     /label=DuetDOWN
                     /note="color: black; sequence: gattatgcggccgtgtacaa"
     primer_bind     2105..2124
                     /label=DuetUP2
                     /note="color: black; sequence: TTGTACACGGCCGCATAATC"
     promoter        2130..2148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    2149..2173
                     /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"
     RBS             2202..2207
                     /note="color: #c0c0c0"
     CDS             2216..2218
                     /codon_start=1
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     CDS             2240..3340
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /note="color: #cc99b2"
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     CDS             3380..3433
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=2¡ÁHA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYAYPYDVPDYA"
     terminator      3500..3547
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     CDS             complement(3767..4426)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /note="color: #ccffcc"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4427..4529)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(5055..5599)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
                     /note="color: #ffff00"
     CDS             complement(5824..6906)
                     /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(6907..6984)
                     /gene="lacI"
                     /label=lacI promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gggatctcga cgctctccct tatgcgactc ctgcattagg aaattaatac gactcactat
       61 aggggaattg tgagcggata acaattcccc tgtagaaata attttgttta actttaataa
      121 ggagatatac catgggcagc agcgcatcac catcatcacc aagccaggga tccatgcaga
      181 agcgaatagc cttgtcgttt ccagaagagg tactagagca tgtgttctcg tttattcagc
      241 tggataagga taggaactca gtctctctgg tgtgcaagtc atggtacgag atcgagcggt
      301 ggtgcaggag gaaagtcttc atcgggaact gctacgccgt gagtccagcg acggtgatta
      361 ggaggttccc gaaagtgaga tccgtggagc ttaaaggaaa acctcacttt gctgacttta
      421 atttggtacc tgacggatgg ggaggttacg tgtatccatg gattgaggcc atgtcttcgt
      481 cttacacgtg gcttgaagag ataaggctga agaggatggt ggtcaccgac gattgcttgg
      541 agctcatagc caagtctttt aagaatttta aggttcttgt gctttcttcc tgcgaaggct
      601 tctccaccga tggtctcgct gctatcgctg ccacttgcag gaatctgaaa gagcttgact
      661 tacgagagag tgatgttgac gacgttagtg gccactggct tagccatttc ccagatacat
      721 acacttcttt ggtatcactc aatatatctt gcttagcatc tgaggtcagt ttctctgctc
      781 tggaaaggct ggtgactagg tgtcccaatc tcaagtctct caagcttaac cgagctgttc
      841 cacttgaaaa attggctact ttacttcaaa gagcacctca attggaggaa ttgggcactg
      901 gtgggtacac tgcagaagtg cgaccagatg tttactctgg tttatctgta gcgctctctg
      961 ggtgcaagga attgaggtgc ttatctggat tttgggatgc tgttcctgcc tatcttccag
     1021 cagtttattc ggtttgcagt cggcttacaa ctttgaatct gagttatgca acagtccaga
     1081 gctatgatct tgtcaagctt ctttgtcaat gccctaaact gcagcgcctc tgggtgcttg
     1141 actacatcga ggatgctggt cttgaggtgc ttgcttcaac ctgcaaggac ctacgcgagc
     1201 tgagagtgtt tccgtccgag ccttttgtca tggaaccaaa tgtggcattg acggaacagg
     1261 ggcttgtctc cgtctccatg ggctgtccaa aactcgagtc ggttctctac ttctgccgtc
     1321 aaatgaccaa tgctgcattg ataaccattg ctaggaaccg tcccaacatg actcgcttcc
     1381 gtttgtgcat cattgagcca aaagccccag actatctgac tctagagcca ctggatattg
     1441 gatttggagc catagtagag cactgcaagg atctccgtcg cctctctcta tctggcctct
     1501 tgaccgacaa ggtttttgaa tacattggga catatgccaa gaagatggaa atgctctcag
     1561 tggcatttgc aggagacagt gacttaggca tgcatcatgt tttgtccggg tgcgatagct
     1621 tgaggaaact agagataagg gactgcccgt ttggagacaa ggcgcttttg gccaatgctt
     1681 caaagctgga gacaatgcga tccctttgga tgtcttcttg ttccgtgagt tttggagcct
     1741 gcaagttact aggacagaag atgccaaagc tgaatgtgga agtcatcgat gaacggggtg
     1801 caccggactc gagaccagag agctgccctg ttgagagagt cttcatatac cgaacagtgg
     1861 ctggtcctcg atttgacatg cctggcttcg tctggaacat ggaccaagac tcaacaatga
     1921 ggttttccag gcaaatcatt actactaacg gattaggatc cgagctcgat atcactagtc
     1981 ctgagcagaa gctcatctcc gaggaggacc tcgagcagaa gctcatctcc gaggaggacc
     2041 tcctgactgc aggtcgacaa gcttgcggcc gcataatgct taagtcgaac agaaagtaat
     2101 cgtattgtac acggccgcat aatcgaaatt aatacgactc actatagggg aattgtgagc
     2161 ggataacaat tccccatctt agtatattag ttaagtataa gaaggagata tacatatggc
     2221 agatctcaat tgggaattca tgaaaatcga agaaggtaaa ctggtaatct ggattaacgg
     2281 cgataaaggc tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat
     2341 taaagtcacc gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac
     2401 tggcgatggc cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc
     2461 tggcctgttg gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac
     2521 ctgggatgcc gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt
     2581 atcgctgatt tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc
     2641 ggcgctggat aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga
     2701 accgtacttc acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatgaaaa
     2761 cggcaagtac gacattaaag acgtgggcgt ggataacgct ggcgcgaaag cgggtctgac
     2821 cttcctggtt gacctgatta aaaacaaaca catgaatgca gacaccgatt actccatcgc
     2881 agaagctgcc tttaataaag gcgaaacagc gatgaccatc aacggcccgt gggcatggtc
     2941 caacatcgac accagcaaag tgaattatgg tgtaacggta ctgccgacct tcaagggtca
     3001 accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt aacgccgcca gtccgaacaa
     3061 agagctggca aaagagttcc tcgaaaacta tctgctgact gatgaaggtc tggaagcggt
     3121 taataaagac aaaccgctgg gtgccgtagc gctgaagtct tacgaggaag agttggcgaa
     3181 agatccacgt attgccgcca ctatggaaaa cgcccagaaa ggtgaaatca tgccgaacat
     3241 cccgcagatg tccgctttct ggtatgccgt gcgtactgcg gtgatcaacg ccgccagcgg
     3301 tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact gaattcgagc ccgggaggcc
     3361 tctcggcgcg cctgcaggtt acccatacga cgtaccagat tacgcttacc catacgacgt
     3421 accagattac gctcgacaag cttgcggccg cataatgctt aagtaattaa cctaggctgc
     3481 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3541 ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     3601 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     3661 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     3721 gcgtagcacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     3781 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     3841 agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     3901 atttgcccat agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     3961 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     4021 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     4081 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     4141 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     4201 ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     4261 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     4321 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     4381 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     4441 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     4501 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
     4561 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
     4621 caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta
     4681 tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca
     4741 gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt
     4801 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
     4861 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
     4921 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
     4981 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
     5041 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
     5101 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tcccctggcg
     5161 gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat
     5221 ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag
     5281 ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat
     5341 cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat
     5401 tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag
     5461 ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag
     5521 aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg
     5581 cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc
     5641 agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta
     5701 attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5761 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5821 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5881 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5941 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     6001 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     6061 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     6121 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     6181 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     6241 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     6301 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     6361 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6421 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6481 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6541 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6601 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6661 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6721 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6781 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6841 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6901 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6961 ggttttgcgc cattcgatgg tgtcc
//
