LOCUS       Exported                8234 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 8234)
  AUTHORS   Broad Institute
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Mar 6, 2021 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8234
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    3..27
                     /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             71..889
                     /codon_start=1
                     /label=AvTnsA
                     /translation="MVRGRREWTQAKYERYIKEGRGRGSGKHYKPWLTIQDVPSKGRSS
                     RTPGWKTNRVHHFFSDHEKRLFYLFEWSDIVTDIREQFPLDNLDLAMSIATDIGIKYPV
                     DTQSGTPHVLTSDFMLTVNRDGKQMQIARTVKPSTELEKKQVNEKFEIERQYYLAKGID
                     WGIITEKEIPRLLAENVEWVHSAYRLEANSEMDTAQLSELSTILKSKLQEEDTTINRVT
                     TALDKEMNINSGTSLYVFKHLIARKEILVDLVAIKLSACPSTKVIEKIIF"
     CDS             912..3140
                     /codon_start=1
                     /label=AvTnsB
                     /translation="MVKDLFVNDLIEWIVYQNTHIVERIIWIDEGYIIAFVIDINAKKG
                     LPEPRKISDILEAITEGIAFKQQQDPWARIIRDENLTDKEKEYRDKAWEIISSLIIEEP
                     SIYYRNFRGSMVKEAVEKYNEERDKDKLVEKTVYGYLRRFWQRGKTINALIPDFINSSG
                     KGKIRGQSGKKRGRPRKYAHDPDIGEGVNVTEEDRKIFRVSISKFYNNPKENYLTTAYE
                     LMVKEYYKEGIRYDENGVQKSILTPTDKIPTITQFKYWYQLEQDDVTKTLTSRKGAKRV
                     ALENRAILGNSKMETIGPGSRYQIDATVADVYLVSKYNRDWIIGRPVIYVIIDVFSRMI
                     TGVYVGLEGPSWIGAMMALANVATDKTLFCQEYNINITENEWPCNQMPDAILGDRGELA
                     GMTVETLIPNLYVRIENAASYRADWKGLVERHFRIIHEYVKPFLPGYVDTDFRQRGARD
                     YRLDGKLDIDQFTEIIIRLIVHHNNKPLTEYERDEAMIADDTPPIPRELWKWGIANRSG
                     RLRTFSEDIVKLNLMPTEKATITAKGIKMRGKEMYYTCDSAKNEQWFEKARSNLLSKSE
                     KSLIVSYDIRKPNFIYLPSSDGRDFEKCFLIDLEGRYANKNFHDIEYLLAYEELQRQKS
                     QGKQLQEKVDLIADIESIVDRAEKMTDASQYHNLSKSKKVSGIRDNRAFEKAKRRENEG
                     FELAITDNQSSFQVAESTAETSEQPESLQPNHLNLLKKKRQERKRGQN"
     CDS             3127..4797
                     /codon_start=1
                     /label=AvTnsC
                     /translation="VDRIESELVKIPNGELAVIARYIDQKLPEYNTNPLIQALPPILSA
                     AEFIDKVAREPNFDEQERELEPHYRFHCIERLSRYFDPQNKTLELQKVICTLIMTGYLS
                     RNLLKPEYANRSRQIYNAIGNGGGKNLENYVNLPTSASGLTLIGPSGIGKTTNFKNILE
                     LYPQIIFHPDYSVYQIVWLRVDCPKTLKGLCIDTFAGIDKLLGTNLLKKFNPARTSEDY
                     LLAQLAQTAHTHHLGVLVIDEIQNLATAKRNRDEILNFLVKLDNTIGIPVIRVGTNEAI
                     SFLQGNLRNARRNTGEGSRMWDRMVNNDEWCYFLEGIWEYQWTKTPVHFSDEIKEVFYD
                     ESQGIIDIAIKLYKMVQWRAISIGDQEIITVDLIRQVAKEGLYLVKPMLDAIRLGDREW
                     MIKYKDIAPLDITEYHNRCVSEIDAKDLTEIKRLAKKQQNQQKVSPKLNYVISELLKLE
                     VESGKAKECAEQIVYSHEDPDIPIPSLVNKAYALALQGCQVNESLAKTSTRKAKLKTKY
                     QENDIRRIVENAKKDKKSAYEDLKLAGIIKNPVEDFINI"
     promoter        4856..4874
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4875..4899
                     /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)."
     terminator      4977..5024
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5254..6069)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(6070..6161)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6179..6814)
                     /direction=LEFT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     CDS             complement(7012..8094)
                     /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(8095..8172)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(8218..8234,1..2)
                     /label=T7 promoter
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atggttaggg gtagaaggga atggactcaa gctaaatatg agcgatatat
      121 aaaagaaggt cgtggccgag gaagtggtaa gcactataaa ccttggctta caatccaaga
      181 tgttccctct aaaggacgtt cttctagaac tcctggttgg aaaacaaatc gagtacatca
      241 tttcttctca gaccatgaaa aacggctatt ttatttgttt gagtggtcag atatagtcac
      301 agatataaga gagcaatttc ctttagacaa tctcgatcta gccatgagta ttgctacaga
      361 cataggtata aagtaccctg tagatacaca aagtggcact cctcacgttt taacaagcga
      421 ctttatgctt acagtcaacc gcgatgggaa gcagatgcag attgcccgga ctgttaagcc
      481 gtcaacagaa ctggagaaaa agcaagttaa tgagaaattc gagatagaaa gacaatatta
      541 cttagcaaag ggtattgact ggggaattat cacagaaaaa gaaattccca ggcttttagc
      601 tgaaaatgtt gagtgggttc attctgctta taggctagaa gcaaattctg aaatggacac
      661 tgcacaactg agtgaattgt caacaatttt aaaatctaag cttcaggaag aagatacaac
      721 tataaataga gtaactactg ctttagacaa agaaatgaat ataaattccg gtacatcact
      781 ttatgtattt aaacatttaa ttgctaggaa agaaatcctt gtggatctag tagcaataaa
      841 attatctgct tgtccgtcaa ctaaagttat tgaaaaaata atattttaga cccacatgat
      901 aggtaaataa tatggttaaa gatttatttg tcaatgattt aatagagtgg attgtatacc
      961 aaaacactca tatagtcgaa agaattattt ggatagatga aggttacatt attgcatttg
     1021 ttatagatat taatgctaag aaaggtctac ctgaaccaag gaaaatatca gatattttag
     1081 aagctataac tgaaggtatt gcttttaagc agcaacaaga tccttgggca agaattattc
     1141 gggatgaaaa tttgacagat aaggaaaagg aatacagaga taaagcttgg gagataattt
     1201 catcattaat tatagaagaa ccatcaatat attatagaaa ttttagagga tcgatggtta
     1261 aagaagcggt tgaaaagtac aacgaagaaa gggataaaga taaattagta gaaaaaaccg
     1321 tctatggtta tttgagaaga ttttggcaga gaggaaaaac tataaatgct ctcatacctg
     1381 attttatcaa ttcatcaggt aagggcaaga ttcgaggaca atcgggcaag aagagaggaa
     1441 gaccaagaaa atatgcacat gacccagata ttggtgaagg tgtaaatgtt acagaagaag
     1501 atagaaagat ttttagagta tctataagca aattttacaa taatccaaaa gaaaactact
     1561 taacaacagc ctatgaatta atggtgaaag aatattacaa ggaaggaatt agatatgacg
     1621 agaatggtgt tcaaaaatct atattaaccc caactgataa aatacctaca attactcaat
     1681 tcaaatactg gtatcaactt gaacaggatg atgttaccaa aactttaaca tctcgaaaag
     1741 gagccaagcg agtagcttta gagaatagag caattttggg aaactctaag atggaaacta
     1801 ttggccctgg ttctagatat caaattgatg ccacagttgc tgatgtatat ttagtttcaa
     1861 aatataaccg tgactggata attggtagac cagttatcta tgtaattatt gatgttttta
     1921 gtagaatgat tacaggtgtt tatgtcggtt tagaaggccc atcttggata ggagcaatga
     1981 tggcactagc caatgtagct acagataaaa ccctgttttg ccaggaatat aacattaata
     2041 ttactgaaaa tgaatggcct tgcaatcaaa tgccagatgc aattttagga gataggggag
     2101 aattagcagg tatgacagta gaaacattaa tccctaatct ttatgttcgt attgaaaatg
     2161 cagcttccta tagagcagat tggaaagggt tagtagaaag acactttcgt ataattcatg
     2221 aatatgtaaa acctttttta cctggttatg tggatacgga ttttaggcaa agaggtgcaa
     2281 gggattatag gcttgatggc aaacttgata tagatcaatt tactgagatt attattcgtt
     2341 taattgttca ccataacaat aaacctctta ctgaatatga aagagatgaa gcaatgattg
     2401 ctgatgatac ccctcctata cctagagagt tgtggaagtg gggtatcgcc aatcgttctg
     2461 gcagactcag aactttttct gaagatattg tcaaattgaa cttgatgcct actgagaaag
     2521 caactattac tgctaaaggc atcaaaatga gaggtaagga gatgtactat acctgtgata
     2581 gtgctaaaaa tgagcaatgg tttgaaaaag ctagaagtaa tctcctttct aagtcagaaa
     2641 aatcattgat tgtatcttat gacatcagaa agccaaactt catttatctt ccatctagtg
     2701 atggtagaga ttttgaaaaa tgttttctta ttgacttaga agggcggtat gctaataaaa
     2761 actttcatga tattgaatat ctgcttgctt atgaagaatt gcaaagacag aaaagccaag
     2821 gtaaacagct acaagaaaag gtagatttaa tagctgatat tgaaagcata gtagatagag
     2881 cagaaaaaat gactgatgct tctcaatatc ataatttaag taagagcaag aaagtttctg
     2941 gaataagaga taatagggct tttgaaaagg ctaaacgacg agagaatgaa ggatttgaat
     3001 tagcaattac agataatcag agcagttttc aggttgcaga gagtacagca gaaacctccg
     3061 aacagccaga atcactacag ccaaaccatc tgaatcttct taagaaaaag cgacaggagc
     3121 gaaaacgtgg acagaattga gagtgaactg gttaaaattc ctaatggaga actagcagtt
     3181 attgccagat atatagatca aaaactgcca gaatacaaca ctaatccatt aattcaggca
     3241 ttacctccta tcctatcagc agcagaattt atagataaag tagccagaga accaaatttt
     3301 gatgaacaag agagagagct agaaccacat tatagatttc attgtattga gaggttatct
     3361 agatattttg accctcaaaa taaaactctt gaattacaaa aagttatttg tacgcttatc
     3421 atgacaggct atctgtcaag gaacctttta aaacctgaat atgcaaatcg ctctaggcaa
     3481 atttataatg caataggtaa tggaggaggt aaaaatttag aaaactatgt taatcttcct
     3541 acatctgcgt ccggcttaac tctaattggc ccatcaggta ttggaaagac tacaaatttc
     3601 aaaaatattc ttgagcttta tccccaaata atttttcatc ctgattatag tgtttaccaa
     3661 atagtttggc tgcgggtaga ttgccctaaa actttaaagg ggttatgtat agatactttt
     3721 gcaggtatcg ataaattgtt aggaactaat ttattaaaga aatttaaccc agcacgcaca
     3781 tctgaagatt atctattggc tcaacttgca caaacagcac acactcatca tctaggagtt
     3841 ttggtgattg atgaaataca aaatttagca actgcaaaaa gaaatagaga cgaaatatta
     3901 aattttttag taaagctcga taatactatt ggaattccag ttattcgtgt aggtacgaat
     3961 gaagccatta gttttcttca aggtaatctt agaaatgcta gaagaaatac tggagaagga
     4021 agtagaatgt gggatagaat ggtaaataat gatgaatggt gttacttttt agaaggaata
     4081 tgggaatacc aatggacaaa aacacctgtt catttctcgg atgaaataaa agaagttttt
     4141 tatgatgaaa gccagggaat tattgatatt gctataaaac tttacaaaat ggttcaatgg
     4201 agagcgattt caataggtga ccaagaaatt attacggttg atctgattcg tcaagtagct
     4261 aaagaaggat tgtatttagt aaagccaatg ctagatgcaa ttaggttagg ggacagagag
     4321 tggatgatta aatataagga tattgctcct ctcgatatta cggaatatca caatagatgt
     4381 gtatctgaga ttgatgctaa agatttgaca gagattaaga gattagctaa aaagcagcag
     4441 aatcagcaga aagtatctcc gaagctcaat tatgtaatta gtgagctgtt gaaattggaa
     4501 gtagaatctg gtaaagcaaa ggaatgtgca gaacaaattg tttatagtca tgaagatcca
     4561 gacattccaa ttccatcttt agttaataag gcatatgctc ttgctttgca aggatgtcaa
     4621 gtaaatgaga gtcttgctaa aacaagtacc aggaaagcta aattaaagac aaagtaccaa
     4681 gagaatgata ttcggcgaat tgtcgagaat gccaagaaag acaaaaaatc tgcatacgag
     4741 gatttgaaat tagcaggtat aatcaaaaat ccagttgaag attttatcaa tatttagctt
     4801 ctaacttaag tcgaacagaa agtaatcgta ttgtacacgg ccgcataatc gaaattaata
     4861 cgactcacta taggggaatt gtgagcggat aacaattccc catcttagta tattagttaa
     4921 gtataagaag gagatatact taattaacct aggctgctgc caccgctgag caataactag
     4981 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa cctcaggcat
     5041 ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac cagcaataga
     5101 cataagcggc tatttaacga ccctgccctg aaccgacgac aagctgacga ccgggtctcc
     5161 gcaagtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5221 cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga gcatcaaatg
     5281 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     5341 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     5401 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     5461 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     5521 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     5581 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcggtc
     5641 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     5701 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     5761 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     5821 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     5881 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     5941 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     6001 taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt cccgttgaat
     6061 atggctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggcatgc tagcgcagaa
     6181 acgtcctaga agatgccagg aggatactta gcagagagac aataaggccg gagcgaagcc
     6241 gtttttccat aggctccgcc cccctgacga acatcacgaa atctgacgct caaatcagtg
     6301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctgatg gctccctctt
     6361 gcgctctcct gttcccgtcc tgcggcgtcc gtgttgtggt ggaggcttta cccaaatcac
     6421 cacgtcccgt tccgtgtaga cagttcgctc caagctgggc tgtgtgcaag aaccccccgt
     6481 tcagcccgac tgctgcgcct tatccggtaa ctatcatctt gagtccaacc cggaaagaca
     6541 cgacaaaacg ccactggcag cagccattgg taactgagaa ttagtggatt tagatatcga
     6601 gagtcttgaa gtggtggcct aacagaggct acactgaaag gacagtattt ggtatctgcg
     6661 ctccactaaa gccagttacc aggttaagca gttccccaac tgacttaacc ttcgatcaaa
     6721 ccgcctcccc aggcggtttt ttcgtttaca gagcaggaga ttacgacgat cgtaaaagga
     6781 tctcaagaag atcctttacg gattcccgac accatcactc tagatttcag tgcaatttat
     6841 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgtaat tctcatgtta
     6901 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     6961 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     7021 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     7081 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     7141 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     7201 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     7261 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     7321 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     7381 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     7441 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     7501 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     7561 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     7621 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     7681 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     7741 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     7801 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     7861 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     7921 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     7981 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     8041 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     8101 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     8161 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaaattaa
     8221 tacgactcac tata
//
