LOCUS       Exported                5770 bp DNA     circular SYN 08-APR-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5770)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 13, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5770
                     /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"
     CDS             195..254
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=2¡ÁMyc
                     /note="color: #cc99b2"
                     /translation="EQKLISEEDLEQKLISEEDL"
     primer_bind     complement(317..336)
                     /label=DuetDOWN
                     /note="color: black; sequence: gattatgcggccgtgtacaa"
     primer_bind     317..336
                     /label=DuetUP2
                     /note="color: black; sequence: TTGTACACGGCCGCATAATC"
     promoter        342..360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    361..385
                     /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             414..419
                     /note="color: #c0c0c0"
     CDS             428..430
                     /codon_start=1
                     /label=ATG
                     /note="color: #993366"
                     /translation="M"
     CDS             452..1552
                     /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"
     primer_bind     1553..1592
                     /label=IAA6 F
                     /note="color: black; sequence: 
                     gaattcgagCCCGGGAGGCCTATGGCAAAGGAAGGTCTAG"
     CDS             1574..2140
                     /codon_start=1
                     /label=AtIAA6
                     /note="color: #00ffff"
                     /translation="MAKEGLALEITELRLGLPGDNYSEISVCGSSKKKKRVLSDMMTSS
                     ALDTENENSVVSSVEDESLPVVKSQAVGWPPVCSYRRKKNNEEASKAIGYVKVSMDGVP
                     YMRKIDLGSSNSYINLVTVLENLFGCLGIGVAKEGKKCEYIIIYEDKDRDWMLVGDVPW
                     QMFKESCKRLRIVKRSDATGFGLQQD"
     primer_bind     complement(2120..2159)
                     /label=IAA6 R
                     /note="color: black; sequence: 
                     caggcgcgccgagAGGCCTATCTTGCTGGAGACCAAAACC"
     CDS             2165..2218
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=2¡ÁHA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYAYPYDVPDYA"
     terminator      2285..2332
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
                     /note="color: #ffffff"
     CDS             complement(2552..3211)
                     /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(3212..3314)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      complement(3840..4384)
                     /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(4609..5691)
                     /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(5692..5769)
                     /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 tccgagctcg
      181 atatcactag tcctgagcag aagctcatct ccgaggagga cctcgagcag aagctcatct
      241 ccgaggagga cctcctgact gcaggtcgac aagcttgcgg ccgcataatg cttaagtcga
      301 acagaaagta atcgtattgt acacggccgc ataatcgaaa ttaatacgac tcactatagg
      361 ggaattgtga gcggataaca attccccatc ttagtatatt agttaagtat aagaaggaga
      421 tatacatatg gcagatctca attgggaatt catgaaaatc gaagaaggta aactggtaat
      481 ctggattaac ggcgataaag gctataacgg tctcgctgaa gtcggtaaga aattcgagaa
      541 agataccgga attaaagtca ccgttgagca tccggataaa ctggaagaga aattcccaca
      601 ggttgcggca actggcgatg gccctgacat tatcttctgg gcacacgacc gctttggtgg
      661 ctacgctcaa tctggcctgt tggctgaaat caccccggac aaagcgttcc aggacaagct
      721 gtatccgttt acctgggatg ccgtacgtta caacggcaag ctgattgctt acccgatcgc
      781 tgttgaagcg ttatcgctga tttataacaa agatctgctg ccgaacccgc caaaaacctg
      841 ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt
      901 caacctgcaa gaaccgtact tcacctggcc gctgattgct gctgacgggg gttatgcgtt
      961 caagtatgaa aacggcaagt acgacattaa agacgtgggc gtggataacg ctggcgcgaa
     1021 agcgggtctg accttcctgg ttgacctgat taaaaacaaa cacatgaatg cagacaccga
     1081 ttactccatc gcagaagctg cctttaataa aggcgaaaca gcgatgacca tcaacggccc
     1141 gtgggcatgg tccaacatcg acaccagcaa agtgaattat ggtgtaacgg tactgccgac
     1201 cttcaagggt caaccatcca aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc
     1261 cagtccgaac aaagagctgg caaaagagtt cctcgaaaac tatctgctga ctgatgaagg
     1321 tctggaagcg gttaataaag acaaaccgct gggtgccgta gcgctgaagt cttacgagga
     1381 agagttggcg aaagatccac gtattgccgc cactatggaa aacgcccaga aaggtgaaat
     1441 catgccgaac atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa
     1501 cgccgccagc ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga ctgaattcga
     1561 gcccgggagg cctatggcaa aggaaggtct agcactcgag atcacagagc ttcgattggg
     1621 tcttccagga gataattata gcgaaatatc agtatgcgga tcgagtaaga agaagaagag
     1681 ggtgctctcg gatatgatga cctcatcagc gttagatact gagaatgaaa acagcgtcgt
     1741 ttcatcagtt gaagatgaat cactgccggt tgtgaagagt caagcggtgg gatggccacc
     1801 tgtgtgttct tacaggagaa agaagaacaa tgaggaagca tcgaaagcta taggctacgt
     1861 gaaagtgagc atggatggtg tgccatacat gaggaagatt gaccttggtt cgagcaacag
     1921 ttatattaat ctagtcacgg ttcttgagaa tctcttcggc tgtcttggca taggagtggc
     1981 gaaggagggt aagaagtgtg aatacattat tatatacgaa gacaaggata gagactggat
     2041 gctcgtcgga gatgtacctt ggcagatgtt taaagaatca tgcaagaggc tgaggatcgt
     2101 gaagagatca gatgcaactg gttttggtct ccagcaagat aggcctctcg gcgcgcctgc
     2161 aggttaccca tacgacgtac cagattacgc ttacccatac gacgtaccag attacgctcg
     2221 acaagcttgc ggccgcataa tgcttaagta attaacctag gctgctgcca ccgctgagca
     2281 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaacc
     2341 tcaggcattt gagaagcaca cggtcacact gcttccggta gtcaataaac cggtaaacca
     2401 gcaatagaca taagcggcta tttaacgacc ctgccctgaa ccgacgaccg ggtcgaattt
     2461 gctttcgaat ttctgccatt catccgctta ttatcactta ttcaggcgta gcaccaggcg
     2521 tttaagggca ccaataactg ccttaaaaaa attacgcccc gccctgccac tcatcgcagt
     2581 actgttgtaa ttcattaagc attctgccga catggaagcc atcacagacg gcatgatgaa
     2641 cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatagtga
     2701 aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca
     2761 cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca
     2821 ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt
     2881 cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac
     2941 aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata cggaactccg
     3001 gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat
     3061 ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac
     3121 attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa
     3181 cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg
     3241 ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc
     3301 ttacgtgccg atcaacgtct cattttcgcc aaaagttggc ccagggcttc ccggtatcaa
     3361 cagggacacc aggatttatt tattctgcga agtgatcttc cgtcacaggt atttattcgg
     3421 cgcaaagtgc gtcgggtgat gctgccaact tactgattta gtgtatgatg gtgtttttga
     3481 ggtgctccag tggcttctgt ttctatcagc tgtccctcct gttcagctac tgacggggtg
     3541 gtgcgtaacg gcaaaagcac cgccggacat cagcgctagc ggagtgtata ctggcttact
     3601 atgttggcac tgatgagggt gtcagtgaag tgcttcatgt ggcaggagaa aaaaggctgc
     3661 accggtgcgt cagcagaata tgtgatacag gatatattcc gcttcctcgc tcactgactc
     3721 gctacgctcg gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt
     3781 cctggaagat gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt
     3841 ttccataggc tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg
     3901 cgaaacccga caggactata aagataccag gcgtttcccc tggcggctcc ctcgtgcgct
     3961 ctcctgttcc tgcctttcgg tttaccggtg tcattccgct gttatggccg cgtttgtctc
     4021 attccacgcc tgacactcag ttccgggtag gcagttcgct ccaagctgga ctgtatgcac
     4081 gaaccccccg ttcagtccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4141 ccggaaagac atgcaaaagc accactggca gcagccactg gtaattgatt tagaggagtt
     4201 agtcttgaag tcatgcgccg gttaaggcta aactgaaagg acaagttttg gtgactgcgc
     4261 tcctccaagc cagttacctc ggttcaaaga gttggtagct cagagaacct tcgaaaaacc
     4321 gccctgcaag gcggtttttt cgttttcaga gcaagagatt acgcgcagac caaaacgatc
     4381 tcaagaagat catcttatta atcagataaa atatttctag atttcagtgc aatttatctc
     4441 ttcaaatgta gcacctgaag tcagccccat acgatataag ttgtaattct catgttagtc
     4501 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     4561 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     4621 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4681 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     4741 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     4801 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4861 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     4921 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     4981 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     5041 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     5101 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     5161 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     5221 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     5281 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5341 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5401 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5461 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5521 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5581 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5641 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     5701 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     5761 gatggtgtcc
//
