LOCUS       Exported                8374 bp ds-DNA     circular SYN 06-MAR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8374)
  AUTHORS   Broad Institute
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Mar 7, 2021 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8374
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        67..85
                     /label=T7 promoter
     protein_bind    86..110
                     /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)."
     CDS             154..2850
                     /codon_start=1
                     /label=PmcTnsAB
                     /translation="MKEQFEQWVARLQISEAGQKIIEKVRSSEPSRRVGGGSKNVSGRY
                     PSRKMGVTIQFESHRVELPIIYQLEHDKDVLEFYDQPPQIKLDYQASNGRRLGVLHTPD
                     LFVIRTNSAGWEECKTEQDLKKLAEKNPNRYFYSEENNQWHSLPGEVYANQFGLYHRIH
                     SDREINWVLQRNLQFLEDYYRSESLVVEEAIAQSLLAIVSSQPGITLAELLNRSTGAKS
                     DDIYTLIIQEQIYIDLNATPLAEPERCLIFRDEQTASAYRLMVEQPSVSLPAISPVVNI
                     VTGATVSWDGKGLNIIHVGETEVILGAENNQLIELKKAIFENLVRQGKITSLQTPEKTA
                     ISTESWQRFHQASPEDQAEALERYKTIEPYLNGHPPENETIPARTIRHWKAKYLIAIQK
                     YGCGYIGLLSHRSVKGNRQRKLPEDTLAIMEKFILEDYETLKQKRMWEVHAALVRACEE
                     AGVIAPSYKAFTKEVQRRTGYEQTKKRQGRRAAYQHESFYWELAITTPRHGDRPFEIGH
                     IDHTELDVELVCSDTGRNLGRPWATFLVDAYSRRLLAVYLTFDSPSYRSCLMVLRICVK
                     RHGRLPQIVVVDNGAEFHSVYFETLLATFECTKKQRPPAKARFGSVCERLFGTSNTQFV
                     HNLLGNTQITRNVRQVTKSVNPKNLAVWTLGLLYEYLCAWAYEVYDTDEHPALFQSPRD
                     AFAAGMAIGGSRVHRMIPYDENFQILTLPTTSEGKAKVQVGRGIKINYIYYWSNSFRDP
                     QIENTSVQIRYDPFNIGIAYAFVRGQWVQCISQYYAELQGRSEKELKLASIELRKHYSN
                     HTQQSKVSAKNLAEFLASVEAQEALLEQRSYDAEVKEVFRVIEGGKATTSRNEEPKLIQ
                     VTFANTDFQADEDEAIVPETLVVYEEF"
     CDS             2853..4004
                     /codon_start=1
                     /label=PmcTnsC
                     /translation="MTKSTGFPLELLTRPATERLAYFENYTVAHPRLKEVYEILMRTIA
                     EPAGASFIFVYGASGVGKTTLRLRVEQKLTELALPKLESDRARVPVVGIEAIAPESRYF
                     NWKEYYTRALITLEEPLIDHKFDYGVRGISRDNFGKINVESKVVAPALRRALENALIHR
                     HPDVFFVDEAQHFGKVASGYKLQDQLDCLKSLANMTGILHCLLGTYELLTFRNLSGQLS
                     RRSVDIHFRRYCADSPEDVQAFKSVLLTFQQHLPLAETPNLVDHWEYFYERTLGCIGTL
                     KDWLKRVLSDALDREATTITLKDLQKRALSVAQCQKMFKEIQEGERQLSETEADVQNLR
                     SALGLGAKPIVLPEETPKTTRPPGKVGKRKPKRDPIGVQQDVS"
     promoter        4056..4074
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4075..4099
                     /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)."
     CDS             4140..5129
                     /codon_start=1
                     /label=PmcTniQ
                     /translation="MTIYSEHWSLNPLEIPQRSRLFSLEPVAVGTPYAESLSSYLHRLA
                     QAHCLTSEKLVMGEIAPLILKDEDKSELLSKNLSHLLGNSDAKPAINGMREMTEKLVTV
                     LEELTMRQDLRFLTLLSWKGMIYDKGLFRNYRAWCPCCCEEWMQKNKTIYEPLSWSFKD
                     VEFCLIHKQRLIEECSHCGARLPVMARLSPAGFCSRCYGWLGQEIKGEEEIEKYRVNIQ
                     GISELIALTPQLGYKPIPIELTRKLQLILLVFEQAIGKDVKLLGDLGGIMESLRIASTT
                     NQSQPYHLVKLIIPVCEKAKISVFQLFGSDFKELGKILFGNFSLELKL"
     terminator      5167..5214
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5335..6126)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(6127..6218)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6266..7004)
                     /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."
     CDS             complement(7214..8296)
                     /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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(8297..8374)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aagcggctat ttaacgaccc tcgggatctc gacgctctcc cttatgcgac tcctgcatta
       61 ggaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc cctgtagaaa
      121 taattttgtt taactttaat aaggagatat accatgaaag agcaatttga acaatgggtt
      181 gctcgccttc aaatatccga ggcagggcaa aaaattattg agaaagttcg ctcatctgag
      241 ccatcccgtc gcgttggtgg cggtagcaag aatgtttcgg gacgctaccc tagtcgcaag
      301 atgggagtca cgattcaatt tgagtctcat cgggttgaac tgccaattat ctatcagtta
      361 gaacatgata aagatgttct agaattctac gatcaaccac cgcaaattaa acttgactat
      421 caagcaagta acggacgtag attaggcgtt ttacacaccc cagatttatt tgtcatccga
      481 accaattcgg ctggttggga agagtgtaaa actgagcagg atttgaaaaa actagcagag
      541 aaaaacccca atcgttactt ttactctgaa gaaaataatc aatggcattc cttgccagga
      601 gaagtttatg ctaaccagtt tgggctttat caccgcattc attcagatcg cgaaatcaat
      661 tgggttctgc aacgtaattt gcaattttta gaagattatt atcgctcaga atctctagta
      721 gtggaagaag cgatcgctca atcattactt gcaattgtgt catctcaacc aggaatcact
      781 ttagcagaat tactcaatcg gtcaacagga gctaaatctg acgatatcta caccttaatt
      841 attcaagagc aaatctatat tgatttaaat gctacaccac tagctgaacc agaaagatgt
      901 ctaatcttcc gtgacgagca aactgctagt gcttatagat taatggttga gcaaccaagt
      961 gtaagtcttc cggctatctc tcctgtagtc aatattgtta ctggagcaac tgttagttgg
     1021 gacggcaaag gtttaaatat catccatgtt ggagaaacag aggttatact tggcgcagaa
     1081 aataatcagt taattgaact caaaaaagct atatttgaga atctggttcg ccaaggaaaa
     1141 atcacaagtc tccaaacacc agaaaaaaca gctattagca cagaatcttg gcagcgattt
     1201 catcaagcta gtcctgaaga ccaagcagaa gcccttgaac gctacaaaac tatcgaacct
     1261 tacttaaacg gacatcctcc agaaaacgaa acaataccag cccgtaccat tcgtcactgg
     1321 aaagctaaat atttaattgc aattcaaaaa tacggttgcg gctacattgg gcttttgtcc
     1381 catcgcagtg tcaaaggtaa tcgacagcgc aagctgcccg aagacacctt agcaattatg
     1441 gagaagttta ttttagaaga ctatgaaact cttaaacaaa aaagaatgtg ggaggttcac
     1501 gcggctttag ttcgagcttg cgaagaagct ggtgtaattg ctccgagcta taaagcattt
     1561 accaaagaag tacaacgccg gactggttac gaacagacta aaaaacgcca aggtcgtcgt
     1621 gccgcttatc agcatgaatc cttctattgg gagcttgcta taactacccc aagacacggc
     1681 gatcgccctt ttgaaattgg gcatatagat catactgagt tagatgtaga gctagtttgc
     1741 tctgatacag gtcgcaatct tggcagacct tgggcaacat ttctcgtaga tgcttattca
     1801 cggcgattgc tggcagttta cctaacgttt gactcaccct catatcgttc ttgcttaatg
     1861 gtgctaagaa tttgtgtcaa gcgtcatggc aggcttcccc aaattgtagt ggtagataac
     1921 ggtgcagagt ttcacagcgt ttactttgaa acactactag ctacttttga atgtacaaaa
     1981 aagcaaagac ccccagccaa agctcgtttt ggttcggttt gcgaacgcct atttggaact
     2041 tccaatactc aatttgtcca taacttactt ggtaatacac aaatcactcg taatgtccgc
     2101 caagtcacta agtcagttaa ccctaaaaat ctagctgtat ggacacttgg tttactttat
     2161 gaatatcttt gtgcttgggc ttacgaagtt tatgatacag acgaacatcc ggcacttttc
     2221 cagagtccga gggatgcgtt tgctgcgggt atggcgattg gaggcagtcg ggttcaccgc
     2281 atgattcctt acgatgaaaa ctttcagatt cttaccctac ctacaacctc agagggaaaa
     2341 gcaaaagttc aagttggacg cggcataaaa attaactaca tatactactg gtctaatagt
     2401 ttccgcgatc ctcaaattga aaatacctct gtgcagataa gatacgaccc gtttaatatt
     2461 ggcatcgctt acgcctttgt tcgaggacag tgggtacagt gtatctctca atattatgca
     2521 gaactccaag gtcgttcaga aaaagaactg aagttagcta gtatcgagtt acgcaaacac
     2581 tattcaaatc atacacaaca atctaaagtt tcggctaaga atttagcaga atttttagca
     2641 tcggtagaag ctcaagaagc tctattagaa cagcgttcct atgatgcaga agttaaggaa
     2701 gtctttaggg taatcgaggg tggaaaagcg acaacaagcc gtaacgaaga accaaaactt
     2761 attcaagtta cttttgccaa taccgatttt caagcggatg aagatgaagc gatcgtccca
     2821 gaaacacttg tagtgtatga ggagttttaa ctatgacaaa atctactgga tttccgcttg
     2881 aactcctcac acggcctgct acagaaaggc ttgcttactt tgaaaattac accgtagcgc
     2941 atcctcgact taaagaagtc tatgaaattc tcatgcgaac gattgccgaa ccagcaggag
     3001 cttcatttat ctttgtttac ggtgctagtg gtgttggtaa aacaactttg cgactccgcg
     3061 ttgagcagaa attaacagaa ctagcattac ctaaattgga aagcgatcgc gctcgtgttc
     3121 ctgttgttgg tattgaagct atagcgccag aatcgagata cttcaactgg aaagaatact
     3181 acacacgcgc cttaattact ttggaagaac cattaatcga tcacaagttt gattacggtg
     3241 tgcgtggtat ctctcgcgat aattttggca aaattaatgt tgagtcgaaa gtagttgcac
     3301 ctgctctacg cagagcttta gaaaatgctc tcatccatcg tcatccagac gtattttttg
     3361 tagatgaagc acaacatttt ggtaaggtgg caagcggata caaactacaa gatcaattag
     3421 actgtctcaa atctttagca aatatgacag gcatcttgca ctgtctatta ggaacatacg
     3481 aactacttac attccgtaat ttgagcggtc aactctcacg tcgtagtgtt gatattcatt
     3541 tccgccgtta ttgtgctgat agcccagaag atgtacaggc ttttaagagt gtgttgttaa
     3601 cttttcaaca gcatctgcca ctagcagaaa ctccaaattt agtcgatcat tgggaatatt
     3661 tttacgaacg tactttagga tgtattggca cactgaaaga ttggctgaag cgtgttctta
     3721 gtgatgcttt ggatcgagaa gcaacaacaa tcactctcaa agatttacaa aaacgtgctt
     3781 tgtcggttgc tcagtgtcag aaaatgttca aggaaattca agagggtgaa cgacaattat
     3841 ctgaaacaga agcagatgtg caaaacttac gctctgcatt aggacttggc gcaaaaccaa
     3901 ttgtgctacc cgaagaaacc ccaaaaacta ctcgtccgcc aggaaaggtt ggcaaacgca
     3961 aacctaaaag agatcccatc ggagtgcagc aagatgttag ctagcttaag tcgaacagaa
     4021 agtaatcgta ttgtacacgg ccgcataatc gaaattaata cgactcacta taggggaatt
     4081 gtgagcggat aacaattccc catcttagta tattagttaa gtataagaag gagatataca
     4141 tgactattta ctcagaacat tggagcctaa atcctctaga aatcccgcaa cggagtcggt
     4201 tattttctct tgaacctgtg gcagtgggaa ctccctatgc agaaagtctt agtagctatc
     4261 tccatcgttt agcgcaggcg cattgtttaa cctctgagaa gctcgttatg ggagaaattg
     4321 cgccacttat tctcaaagat gaggataaat ctgagctatt gtcaaaaaat ctgagtcacc
     4381 tattgggaaa tagcgacgct aaaccggcta ttaacggaat gcgagaaatg acagaaaagt
     4441 tagtcactgt cttagaagag ttaacaatgc gtcaggattt acgctttttg acccttttaa
     4501 gctggaaagg aatgatttat gacaagggat tatttcggaa ctatcgcgct tggtgtcctt
     4561 gttgttgtga agagtggatg cagaagaaca aaactatcta tgaaccatta tcatggtcat
     4621 ttaaggacgt agaattttgt ttgattcaca aacagcgatt aatagaagaa tgttctcatt
     4681 gtggtgcccg tttacctgtg atggctaggc tttcacctgc gggattttgt tctcgttgtt
     4741 acgggtggtt aggacaggaa attaaaggcg aagaagaaat agagaaatat cgagttaata
     4801 ttcaaggaat tagcgaatta attgcactta ctccccaatt aggttataaa cccatcccga
     4861 tcgagttaac ccgaaaatta caattgattt tgttagtgtt tgaacaagca attggaaagg
     4921 atgtgaagtt attgggagat ttaggaggaa tcatggaatc gttgaggata gcctcaacaa
     4981 caaatcaaag tcaaccttat catttggtga agttgattat tcctgtgtgc gagaaggcga
     5041 agattagtgt attccagttg tttgggtcag attttaagga attgggcaag atattgttcg
     5101 ggaattttag tctggaatta aagttatagt taattaacct aggctgctgc caccgctgag
     5161 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     5221 cctcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtgccctg aaccgacgac
     5281 cgggtcatcg tggccggatc ttgcggcccc tcggcttgaa cgaattgtta gacattattt
     5341 gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca actgatctgc
     5401 gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt caagtatgac
     5461 gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat ccttcggcgc
     5521 gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta catttcgctc
     5581 atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta gcgcctcaaa
     5641 tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta ccaaggcaac
     5701 gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg tggctggctc
     5761 gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt cgcgcttagc
     5821 tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta cagcgcggag
     5881 aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca aagctcgccg
     5941 cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac tgtgtggctt
     6001 caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg gttcgagatg
     6061 gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga tcaccgcttc
     6121 cctcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6181 cggatacata tttgaatgta tttagaaaaa taaacaaata gctagctcac tcggtcgcta
     6241 cgctccgggc gtgagactgc ggcgggcgct gcggacacat acaaagttac ccacagattc
     6301 cgtggataag caggggacta acatgtgagg caaaacagca gggccgcgcc ggtggcgttt
     6361 ttccataggc tccgccctcc tgccagagtt cacataaaca gacgcttttc cggtgcatct
     6421 gtgggagccg tgaggctcaa ccatgaatct gacagtacgg gcgaaacccg acaggactta
     6481 aagatcccca ccgtttccgg cgggtcgctc cctcttgcgc tctcctgttc cgaccctgcc
     6541 gtttaccgga tacctgttcc gcctttctcc cttacgggaa gtgtggcgct ttctcatagc
     6601 tcacacactg gtatctcggc tcggtgtagg tcgttcgctc caagctgggc tgtaagcaag
     6661 aactccccgt tcagcccgac tgctgcgcct tatccggtaa ctgttcactt gagtccaacc
     6721 cggaaaagca cggtaaaacg ccactggcag cagccattgg taactgggag ttcgcagagg
     6781 atttgtttag ctaaacacgc ggttgctctt gaagtgtgcg ccaaagtccg gctacactgg
     6841 aaggacagat ttggttgctg tgctctgcga aagccagtta ccacggttaa gcagttcccc
     6901 aactgactta accttcgatc aaaccacctc cccaggtggt tttttcgttt acagggcaaa
     6961 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct actgaaccgc
     7021 tctagatttc agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat
     7081 ataagttgta attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     7141 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     7201 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     7261 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     7321 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     7381 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     7441 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     7501 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     7561 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     7621 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     7681 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     7741 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     7801 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     7861 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     7921 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     7981 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     8041 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     8101 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     8161 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     8221 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     8281 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     8341 taccgcgaaa ggttttgcgc cattcgatgg tgtc
//
