LOCUS       40924_43763        8158 bp DNA     circular SYN 15-JUN-2019
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8158)
  AUTHORS   xsan
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8158)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /label=lac operator
                     /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)."
     RBS             59..81
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             88..780
                     /codon_start=1
                     /label=TnsA
                     /note="accession number: WP_000202732.1"
                     /translation="MATSLPTPSAITTSALEYAFHTPARNLTKSRGKNIHRYVSVKMSK
                     RITVESTLECDACYHFDFEPSIVRFCAQPIRFLYYLNGQSHSYVPDFLVQFDTNEFVLY
                     EVKSAYAKNKPDFDVEWEAKVKAATELGLELELVEESDIRDTVVLNNLKRMHRYASKDE
                     LNNVHNSLLKIIKYNGAQSARCLGEQLGLKGRTVLPILCDLLSRCLLDTRLDKPLSLES
                     RFELASYG"
     CDS             2166..2177
                     /codon_start=1
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
                     /translation="IEGR"
     CDS             2596..3588
                     /codon_start=1
                     /label=TnsC(WP_002024152.1)
                     /translation="MSETREARISRAKRAFVSTPSVRKILSYMDRCRDLSDLESEPTCM
                     MVYGASGVGKTTVIKKYLNQNRRESEAGGDIIPVLHIELPDNAKPVDAARELLVEMGDP
                     LALYETDLARLTKRLTELIPAVGVKLIIIDEFQHLVEERSNRVLTQVGNWLKMILNKTK
                     CPIVIFGMPYSKVVLQANSQLHGRFSIQVELRPFSYQGGRGVFKTFLEYLDKALPFEKQ
                     AGLANESLQKKLYAFSQGNMRSLRNLIYQASIEAIDNQHETITEEDFVFASKLTSGDKP
                     NSWKNPFEEGVEVTEDMLRPPPKDIGWEDYLRHSTPRVSKPGRNKNFFE"
     promoter        3615..3633
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3634..3658
                     /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             3701..4885
                     /codon_start=1
                     /label=TniQ(WP_000479715.1)
                     /translation="MFLQRPKPYSDESLESFFIRVANKNGYGDVHRFLEATKRFLQDID
                     HNGYQTFPTDITRINPYSAKNSSSARTASFLKLAQLTFNEPPELLGLAINRTNMKYSPS
                     TSAVVRGAEVFPRSLLRTHSIPCCPLCLRENGYASYLWHFQGYEYCHSHNVPLITTCSC
                     GKEFDYRVSGLKGICCKCKEPITLTSRENGHEAACTVSNWLAGHESKPLPNLPKSYRWG
                     LVHWWMGIKDSEFDHFSFVQFFSNWPRSFHSIIEDEVEFNLEHAVVSTSELRLKDLLGR
                     LFFGSIRLPERNLQHNIILGELLCYLENRLWQDKGLIANLKMNALEATVMLNCSLDQIA
                     SMVEQRILKPNRKSKPNSPLDVTDYLFHFGDIFCLWLAEFQSDEFNRSFYVSRW"
     terminator      4918..4965
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(5198..6010)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     promoter        complement(6011..6102)
                     /label=AmpR promoter
     rep_origin      complement(6120..6755)
                     /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."
     protein_bind    complement(6919..6940)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6956..8035)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(8036..8113)
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg gcgacaagtt tacctacgcc ctcagcaatt
      121 acgacttcgg cgttagagta tgcattccat actcccgctc gcaatctaac gaaatctcgc
      181 ggaaaaaaca ttcatcgtta tgtcagtgta aagatgagta agaggattac ggtagaatct
      241 actctagagt gtgatgcctg ctatcacttt gattttgagc caagtattgt tcgcttttgc
      301 gctcaaccga ttcgattttt atattatctc aatggtcagt ctcactccta tgttcctgac
      361 tttctagttc aatttgatac caacgagttt gttctatatg aagtaaagtc agcttatgct
      421 aagaacaaac ctgattttga tgttgaatgg gaggcgaaag taaaagcagc aactgaacta
      481 gggctagaat tggagcttgt tgaagagagt gatattaggg atacggttgt attaaataat
      541 cttaagcgca tgcatcgtta tgcttcgaaa gatgagctga ataacgtaca taactctctc
      601 ttaaaaataa taaagtacaa tggcgcccaa tctgcaagat gcttgggaga acagttgggt
      661 ttaaaaggcc gaactgtttt accaattttg tgcgatttgc tgtcaaggtg tttactcgat
      721 acacgtttgg ataagcctct atctcttgaa tctcgatttg agttggccag ttatggctaa
      781 gaaagggttc tcaagtttcc atagaaaagc agtctcttcc caagatacgc ttgaatcgat
      841 agagcttgtc tctagcgcta attgtctaga aagtgttacg tatcaagata tatcagcatt
      901 tcccgaaaca attgcggtag agattaattt ccgattaagc attcttcgct ttttagcgcg
      961 aaagtgcgaa accattgtgg ccaaatcaat tgaaccacat cgtgtagagc tacagcaaaa
     1021 ctatagtaga aaaataccca gtgcaataac gatatatcga tggtggcttg cttttcgaaa
     1081 atcagactac aaccccatta gcttagcacc taatatcaag gatagaggta atagagagac
     1141 aaaagtgtca acagttgttg attctattat ggaacaggca gttgaaagag ttatatctgg
     1201 acgaaaagtc aatgttagct ctgcatataa acgtgttcga cgaaaagttc gtcaatacaa
     1261 tctaactcat ggaacgaaat acacgtatcc taagtacgaa tctgtaagaa agcgagtaaa
     1321 aaagaaaacc ccatttgagt tattagccgc agggaaaggg gagagagtag ctaagagaga
     1381 gtttcgccga atgggaaaaa agatcctcac gtctagcgtg ctagagaggg ttgaaataga
     1441 tcacactgtc gttgaccttt ttgcagtaca cgaagagtat cgaatcccat tgggccgacc
     1501 ttggcttact caattggttg attgttacag taaagctgta atcggttttt atttaggttt
     1561 cgagcctcct agctatgtgt cggtttccct tgcacttaag aatgcaatac aacgcaaaga
     1621 tgacttaatc tcctcgtatg aatcgatcga gaatgaatgg ctatgttatg gcatcccaga
     1681 cctactcgta actgataatg gtaaagagtt tttgtcgaaa gcatttgatc aagcatgtga
     1741 atcactattg atcaatgtgc atcaaaataa agttgagacg cccgacaaca aacctcatgt
     1801 tgaacgtaac tacgggacta ttaatacttc tctgttagac gatttacctg ggaaatcctt
     1861 cagccagtac cttcaaagag aagggtacga ctctgtggga gaagctaccc ttacactcaa
     1921 tgagattaga gaaatttact taatttggtt ggtggatatt tatcataaaa aacccaatca
     1981 gagaggcact aattgtccta atgttgcgtg gaaaaagggt tgtcaagaat gggaaccaga
     2041 ggagttctct ggttctaaag acgaattaga ctttaaattt gctattgttg attacaaaca
     2101 acttactaaa gtagggataa ctgtctacaa agaactgagt tatagcaatg accgtttagc
     2161 tgaatataga gggaagaaag gaaaccataa agttcagttc aagtataacc ctgagtgtat
     2221 ggcagttatt tgggtgttgg atgaggatat gaatgagtac tttacagtta atgcgattga
     2281 ctacgaatat gcaagtagag tatcactttg gcaacataaa tataacatga aatatcaagc
     2341 agaactaaat tcagcagaat atgatgagga caaggaaatt gatgcagaaa taaaaattga
     2401 agaaatcgca gatcgttcaa ttgttaagac taacaaaatc agagctcgga ggcgtggcgc
     2461 taggcatcaa gagaatagcg caagggctaa gtcaatcagt aatgcgaacc cggcctcgat
     2521 acaaaaacat gaagatgaaa tcgttagtgc agataatgac gattgggata ttgattatgt
     2581 ctgagaaatc gtcaaatgag tgaaacgcgt gaggctcgta tatcaagagc taaaagggca
     2641 tttgtatcca caccgagcgt taggaaaatc ttgagttaca tggatagatg tagagatcta
     2701 tcagacctag agtctgagcc tacatgcatg atggtctatg gtgcttcggg tgtaggtaaa
     2761 acgaccgtca tcaagaaata cttaaatcaa aacagaagag agtccgaagc cgggggcgat
     2821 ataataccgg ttttgcatat tgagttgcca gacaatgcga agccagtaga tgcagcaagg
     2881 gaattgctgg ttgaaatggg tgacccgcta gcactttatg aaactgactt agctagattg
     2941 acgaaaagac tgactgaatt aatccctgcg gtcggcgtga agctgattat tatcgatgag
     3001 ttccaacatt tggtggaaga aaggtcaaat cgggttctta cccaagtagg taattggcta
     3061 aaaatgatac ttaacaaaac gaaatgtcca attgttatat ttggtatgcc atactcaaaa
     3121 gttgtactgc aagcaaactc gcaacttcac gggcgatttt ccattcaggt tgaactgcgc
     3181 ccctttagct accagggagg tagaggtgta tttaaaactt ttttggaata ccttgataaa
     3241 gccctacctt ttgaaaaaca ggctggctta gccaacgaaa gtttgcagaa aaaattgtat
     3301 gcattctctc agggaaacat gcgttcgttg agaaacctta tttatcaagc atctatcgaa
     3361 gcaattgata atcagcatga gacgataacc gaagaagatt tcgtttttgc atcgaagttg
     3421 acatcgggcg ataaacccaa ctcatggaaa aatccttttg aggagggtgt tgaggtaaca
     3481 gaagatatgt tacgaccgcc accaaaagat attggttggg aggactattt gagacattca
     3541 accccgagag tgagtaaacc aggtagaaat aaaaactttt tcgaataacc taggcacggc
     3601 cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
     3661 atcttagtat attagttaag tataagaagg agatatacat atgtttttgc aaagacctaa
     3721 accttacagc gatgaaagtt tagaaagttt ctttatccga gtggctaaca aaaatggcta
     3781 cggtgatgtc catcgcttcc tagaagccac taaacgattc cttcaagaca ttgaccataa
     3841 tggctatcaa acctttccga ctgatataac tcggataaac ccatactcag ctaaaaacag
     3901 ttccagcgca cgaactgcgt cattcctgaa acttgcacaa ttgacattta atgaaccgcc
     3961 agagctactt gggttggcaa ttaacagaac aaacatgaaa tactcgccgt caactagcgc
     4021 ggttgttaga ggtgcagaag tctttcctcg cagtttacta cggacgcact ccatcccctg
     4081 ctgtcctttg tgtctgcgag aaaatggcta cgcctcctac ctttggcact ttcaggggta
     4141 cgaatactgc cacagccata acgtaccttt aattaccact tgtagctgtg gtaaggagtt
     4201 tgactaccga gtatctgggt taaagggcat ttgctgcaaa tgcaaggagc ctatcacctt
     4261 aaccagcagg gagaacggtc atgaggcagc gtgtactgtt tcaaactggc ttgctggcca
     4321 tgaatctaaa cctctgccaa atcttcctaa aagctaccga tggggtttag ttcattggtg
     4381 gatgggtatt aaagatagcg agttcgatca cttttcgttc gttcaatttt tctcaaactg
     4441 gccaaggtca ttccactcga taatcgaaga tgaagtagag ttcaaccttg agcatgctgt
     4501 tgtcagcacg tctgaattac gactaaaaga tcttcttggt cgattgtttt tcggttcaat
     4561 tcggttacct gagcggaatc ttcaacacaa tatcatcctt ggtgagcttc tctgctattt
     4621 agaaaatcgt ttatggcaag acaagggatt aatcgccaac ctcaaaatga acgcgttaga
     4681 ggcgactgta atgttaaatt gtagcctcga tcagattgca tcaatggttg aacaacgcat
     4741 cttgaagcca aatcgaaaaa gcaagcccaa cagccctctt gatgttaccg attatctatt
     4801 tcatttcggc gatattttct gtctttggtt agctgagttc caaagcgatg agtttaaccg
     4861 ttcgttttat gtgtcgaggt ggtaaatgcc taggctgctg ccaccgctga gcaataacta
     4921 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa acctcaggca
     4981 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     5041 acataagcgg ctatttaacg accctgccct gaaccgacga caagctgacg accgggtctc
     5101 cgcaagtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5161 acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat
     5221 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
     5281 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
     5341 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
     5401 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt
     5461 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
     5521 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcggt
     5581 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
     5641 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
     5701 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
     5761 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
     5821 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat
     5881 acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
     5941 ataaatcagc atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa
     6001 tatggctcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6061 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggcatg ctagcgcaga
     6121 aacgtcctag aagatgccag gaggatactt agcagagaga caataaggcc ggagcgaagc
     6181 cgtttttcca taggctccgc ccccctgacg aacatcacga aatctgacgc tcaaatcagt
     6241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgat ggctccctct
     6301 tgcgctctcc tgttcccgtc ctgcggcgtc cgtgttgtgg tggaggcttt acccaaatca
     6361 ccacgtcccg ttccgtgtag acagttcgct ccaagctggg ctgtgtgcaa gaaccccccg
     6421 ttcagcccga ctgctgcgcc ttatccggta actatcatct tgagtccaac ccggaaagac
     6481 acgacaaaac gccactggca gcagccattg gtaactgaga attagtggat ttagatatcg
     6541 agagtcttga agtggtggcc taacagaggc tacactgaaa ggacagtatt tggtatctgc
     6601 gctccactaa agccagttac caggttaagc agttccccaa ctgacttaac cttcgatcaa
     6661 accgcctccc caggcggttt tttcgtttac agagcaggag attacgacga tcgtaaaagg
     6721 atctcaagaa gatcctttac ggattcccga caccatcact ctagatttca gtgcaattta
     6781 tctcttcaaa tgtagcacct gaagtcagcc ccatacgata taagttgtaa ttctcatgtt
     6841 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     6901 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     6961 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7021 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     7081 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     7141 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     7201 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     7261 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     7321 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     7381 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     7441 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     7501 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     7561 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     7621 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     7681 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     7741 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     7801 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     7861 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     7921 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     7981 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     8041 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     8101 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaaat
//
