LOCUS       40924_44549        7716 bp DNA     circular SYN 13-MAY-2021
DEFINITION  IPTG-inducible expression of Transposon Tn5 fused to Mxe Intein and 
            Chitin-binding domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7716)
  AUTHORS   Picelli S, Bjorklund AK, Reinius B, Sagasser S, Winberg G, Sandberg 
            R
  TITLE     Tn5 transposase and tagmentation procedures for massively-scaled 
            sequencing projects.
  JOURNAL   Genome Res. 2014 Jul 30. pii: gr.177881.114.
  PUBMED    25079858
REFERENCE   2  (bases 1 to 7716)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7716)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome Res.
            2014 Jul 30. pii: gr.177881.114."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(145..333)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(856..877)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(893..1972)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(1973..2050)
                     /label=lacI promoter
     terminator      2203..2289
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2386..2472
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(2572..2615)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        2790..2808
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2809..2833
                     /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)."
     CDS             2877..4304
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     CDS             4305..4898
                     /codon_start=1
                     /label=Mxe GyrA intein
                     /note="modified GyrA intein (Southworth et al., 1999)"
                     /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
                     LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
                     IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
                     AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
     CDS             4929..5084
                     /codon_start=1
                     /label=CBD
                     /note="chitin binding domain from chitinase A1 (Watanabe et
                     al., 1994)"
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     terminator      5163..5210
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5270..5374
                     /label=AmpR promoter
     CDS             5375..6232
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(6277..6790)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      6901..7489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7643..7660
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(7682..7701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
ORIGIN
        1 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
       61 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
      121 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
      181 gctgcggtaa agctcatcag cgtggtcgtg cagcgattca cagatgtctg cctgttcatc
      241 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
      301 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
      361 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
      421 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
      481 gcgggaccag agaaaaatca ctcagggtca atgccagccg aacgccagca agacgtagcc
      541 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
      601 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
      661 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
      721 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
      781 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
      841 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
      901 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
      961 aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag acgggcaaca
     1021 gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc acgctggttt
     1081 gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg cgggatataa catgagctgt
     1141 cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc ccggactcgg
     1201 taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc gcagtgggaa
     1261 cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca ctccagtcgc
     1321 cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc cagccagcca
     1381 gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt tgctggtgac
     1441 ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag aaaataatac
     1501 tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta gtgcaggcag
     1561 cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc ccactgacgc
     1621 gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt cgttctacca
     1681 tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc gcgacaattt
     1741 gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac gactgtttgc
     1801 ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc gccgcttcca
     1861 ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg gaaacggtct
     1921 gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc acattcacca
     1981 ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt
     2041 cgatggtgtc cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc
     2101 agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg ccggcatgcc
     2161 gccctttcgt cttcaagaat taattcccaa ttccccaggc atcaaataaa acgaaaggct
     2221 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     2281 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg
     2341 gcaggacgcc cgccataaac tgccaggaat taattcccca ggcatcaaat aaaacgaaag
     2401 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     2461 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     2521 cgggcaggac gcccgccata aactgccagg aattaattcc ccaggcatca aataaaacga
     2581 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     2641 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     2701 tggcgggcag gacgcccgcc ataaactgcc aggaattggg gatcggaatt aattcccggt
     2761 ttaaaccggg gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
     2821 ggataacaat tcccctctag aaataatttt gttaacttta agaaggagat atacatatga
     2881 ttaccagtgc actgcatcgt gcggcggatt gggcgaaaag cgtgttttct agtgctgcgc
     2941 tgggtgatcc gcgtcgtacc gcgcgtctgg tgaatgttgc ggcgcaactg gccaaatata
     3001 gcggcaaaag cattaccatt agcagcgaag gcagcaaagc catgcaggaa ggcgcgtatc
     3061 gttttattcg taatccgaac gtgagcgcgg aagcgattcg taaagcgggt gccatgcaga
     3121 ccgtgaaact ggcccaggaa tttccggaac tgctggcaat tgaagatacc acctctctga
     3181 gctatcgtca tcaggtggcg gaagaactgg gcaaactggg tagcattcag gataaaagcc
     3241 gtggttggtg ggtgcatagc gtgctgctgc tggaagcgac cacctttcgt accgtgggcc
     3301 tgctgcatca agaatggtgg atgcgtccgg atgatccggc ggatgcggat gaaaaagaaa
     3361 gcggcaaatg gctggccgct gctgcaactt cgcgtctgag aatgggcagc atgatgagca
     3421 acgtgattgc ggtgtgcgat cgtgaagcgg atattcatgc gtatctgcaa gataaactgg
     3481 cccataacga acgttttgtg gtgcgtagca aacatccgcg taaagatgtg gaaagcggcc
     3541 tgtatctgta tgatcacctg aaaaaccagc cggaactggg cggctatcag attagcattc
     3601 cgcagaaagg cgtggtggat aaacgtggca aacgtaaaaa ccgtccggcg cgtaaagcga
     3661 gcctgagcct gcgtagcggc cgtattaccc tgaaacaggg caacattacc ctgaacgcgg
     3721 tgctggccga agaaattaat ccgccgaaag gcgaaacccc gctgaaatgg ctgctgctga
     3781 ccagcgagcc ggtggaaagt ctggcccaag cgctgcgtgt gattgatatt tatacccatc
     3841 gttggcgcat tgaagaattt cacaaagcgt ggaaaacggg tgcgggtgcg gaacgtcagc
     3901 gtatggaaga accggataac ctggaacgta tggtgagcat tctgagcttt gtggcggtgc
     3961 gtctgctgca actgcgtgaa tcttttactc cgccgcaagc actgcgtgcg cagggcctgc
     4021 tgaaagaagc ggaacacgtt gaaagccaga gcgcggaaac cgtgctgacc ccggatgaat
     4081 gccaactgct gggctatctg gataaaggca aacgcaaacg caaagaaaaa gcgggcagcc
     4141 tgcaatgggc gtatatggcg attgcgcgtc tgggcggctt tatggatagc aaacgtaccg
     4201 gcattgcgag ctggggtgcg ctgtgggaag gttgggaagc gctgcaaagc aaactggatg
     4261 gctttctggc cgcgaaagac ctgatggcgc agggcattaa aatctgcatc acgggagatg
     4321 cactagttgc cctacccgag ggcgagtcgg tacgcatcgc cgacatcgtg ccgggtgcgc
     4381 ggcccaacag tgacaacgcc atcgacctga aagtccttga ccggcatggc aatcccgtgc
     4441 tcgccgaccg gctgttccac tccggcgagc atccggtgta cacggtgcgt acggtcgaag
     4501 gtctgcgtgt gacgggcacc gcgaaccacc cgttgttgtg tttggtcgac gtcgccgggg
     4561 tgccgaccct gctgtggaag ctgatcgacg aaatcaagcc gggcgattac gcggtgattc
     4621 aacgcagcgc attcagcgtc gactgtgcag gttttgcccg cgggaaaccc gaatttgcgc
     4681 ccacaaccta cacagtcggc gtccctggac tggtgcgttt cttggaagca caccaccgag
     4741 acccggacgc ccaagctatc gccgacgagc tgaccgacgg gcggttctac tacgcgaaag
     4801 tcgccagtgt caccgacgcc ggcgtgcagc cggtgtatag ccttcgtgtc gacacggcag
     4861 accacgcgtt tatcacgaac gggttcgtca gccacgctac tggcctcacc ggtctgaact
     4921 caggcctcac gacaaatcct ggtgtatccg cttggcaggt caacacagct tatactgcgg
     4981 gacaattggt cacatataac ggcaagacgt ataaatgttt gcagccccac acctccttgg
     5041 caggatggga accatccaac gttcctgcct tgtggcagct tcaatgactg caggaagggg
     5101 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     5161 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     5221 gaactatatc cggataacta cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     5281 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     5341 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     5401 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5461 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5521 caacagcggt aagatccttg agagttttcg ccccgaagaa cgtttcccaa tgatgagcac
     5581 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     5641 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5701 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5761 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5821 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5881 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5941 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6001 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6061 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6121 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6181 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6241 agaccaagtt tactcatata tactttagat tgatttaccc cggttgataa tcagaaaagc
     6301 cccaaaaaca ggaagattgt ataagcaaat atttaaattg taaacgttaa tattttgtta
     6361 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     6421 aaaatccctt ataaatcaaa agaatagccc gagatagggt tgagtgttgt tccagtttgg
     6481 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     6541 cagggcgatg gcccactacg tgaaccatca cccaaatcaa gttttttggg gtcgaggtgc
     6601 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     6661 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     6721 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6781 cagggcgcgt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6841 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6901 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6961 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7021 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     7081 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7141 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7201 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7261 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7321 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7381 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7441 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7501 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7561 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     7621 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagctatgg tgcactctca
     7681 gtacaatctg ctctgatgcc gcatagttaa gccagt
//
