LOCUS       Exported                7899 bp DNA     circular SYN 20-JUL-2025
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 7899)
  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 7899)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7899)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7899)
  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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2199..2381
                     /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      2386..2472
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2569..2655
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(2755..2798)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        2973..2991
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2992..3016
                     /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             3060..4487
                     /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             4488..5081
                     /codon_start=1
                     /label=Mxe GyrA intein
                     /note="modified GyrA intein (Southworth et al., 1999)"
                     /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
                     LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
                     IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
                     AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
     CDS             5112..5267
                     /codon_start=1
                     /label=CBD
                     /note="chitin binding domain from chitinase A1 (Watanabe et
                     al., 1994)"
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     terminator      5346..5393
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        5453..5557
                     /label=AmpR promoter
     CDS             5558..6415
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      complement(6460..6973)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      7084..7672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7826..7843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(7865..7884)
                     /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 aggaattaat tccccaggca tcaaataaaa
     2761 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2821 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     2881 gggtggcggg caggacgccc gccataaact gccaggaatt ggggatcgga attaattccc
     2941 ggtttaaacc ggggatctcg atcccgcgaa attaatacga ctcactatag gggaattgtg
     3001 agcggataac aattcccctc tagaaataat tttgttaact ttaagaagga gatatacata
     3061 tgattaccag tgcactgcat cgtgcggcgg attgggcgaa aagcgtgttt tctagtgctg
     3121 cgctgggtga tccgcgtcgt accgcgcgtc tggtgaatgt tgcggcgcaa ctggccaaat
     3181 atagcggcaa aagcattacc attagcagcg aaggcagcaa agccatgcag gaaggcgcgt
     3241 atcgttttat tcgtaatccg aacgtgagcg cggaagcgat tcgtaaagcg ggtgccatgc
     3301 agaccgtgaa actggcccag gaatttccgg aactgctggc aattgaagat accacctctc
     3361 tgagctatcg tcatcaggtg gcggaagaac tgggcaaact gggtagcatt caggataaaa
     3421 gccgtggttg gtgggtgcat agcgtgctgc tgctggaagc gaccaccttt cgtaccgtgg
     3481 gcctgctgca tcaagaatgg tggatgcgtc cggatgatcc ggcggatgcg gatgaaaaag
     3541 aaagcggcaa atggctggcc gctgctgcaa cttcgcgtct gagaatgggc agcatgatga
     3601 gcaacgtgat tgcggtgtgc gatcgtgaag cggatattca tgcgtatctg caagataaac
     3661 tggcccataa cgaacgtttt gtggtgcgta gcaaacatcc gcgtaaagat gtggaaagcg
     3721 gcctgtatct gtatgatcac ctgaaaaacc agccggaact gggcggctat cagattagca
     3781 ttccgcagaa aggcgtggtg gataaacgtg gcaaacgtaa aaaccgtccg gcgcgtaaag
     3841 cgagcctgag cctgcgtagc ggccgtatta ccctgaaaca gggcaacatt accctgaacg
     3901 cggtgctggc cgaagaaatt aatccgccga aaggcgaaac cccgctgaaa tggctgctgc
     3961 tgaccagcga gccggtggaa agtctggccc aagcgctgcg tgtgattgat atttataccc
     4021 atcgttggcg cattgaagaa tttcacaaag cgtggaaaac gggtgcgggt gcggaacgtc
     4081 agcgtatgga agaaccggat aacctggaac gtatggtgag cattctgagc tttgtggcgg
     4141 tgcgtctgct gcaactgcgt gaatctttta ctccgccgca agcactgcgt gcgcagggcc
     4201 tgctgaaaga agcggaacac gttgaaagcc agagcgcgga aaccgtgctg accccggatg
     4261 aatgccaact gctgggctat ctggataaag gcaaacgcaa acgcaaagaa aaagcgggca
     4321 gcctgcaatg ggcgtatatg gcgattgcgc gtctgggcgg ctttatggat agcaaacgta
     4381 ccggcattgc gagctggggt gcgctgtggg aaggttggga agcgctgcaa agcaaactgg
     4441 atggctttct ggccgcgaaa gacctgatgg cgcagggcat taaaatctgc atcacgggag
     4501 atgcactagt tgccctaccc gagggcgagt cggtacgcat cgccgacatc gtgccgggtg
     4561 cgcggcccaa cagtgacaac gccatcgacc tgaaagtcct tgaccggcat ggcaatcccg
     4621 tgctcgccga ccggctgttc cactccggcg agcatccggt gtacacggtg cgtacggtcg
     4681 aaggtctgcg tgtgacgggc accgcgaacc acccgttgtt gtgtttggtc gacgtcgccg
     4741 gggtgccgac cctgctgtgg aagctgatcg acgaaatcaa gccgggcgat tacgcggtga
     4801 ttcaacgcag cgcattcagc gtcgactgtg caggttttgc ccgcgggaaa cccgaatttg
     4861 cgcccacaac ctacacagtc ggcgtccctg gactggtgcg tttcttggaa gcacaccacc
     4921 gagacccgga cgcccaagct atcgccgacg agctgaccga cgggcggttc tactacgcga
     4981 aagtcgccag tgtcaccgac gccggcgtgc agccggtgta tagccttcgt gtcgacacgg
     5041 cagaccacgc gtttatcacg aacgggttcg tcagccacgc tactggcctc accggtctga
     5101 actcaggcct cacgacaaat cctggtgtat ccgcttggca ggtcaacaca gcttatactg
     5161 cgggacaatt ggtcacatat aacggcaaga cgtataaatg tttgcagccc cacacctcct
     5221 tggcaggatg ggaaccatcc aacgttcctg ccttgtggca gcttcaatga ctgcaggaag
     5281 gggatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     5341 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     5401 gaggaactat atccggataa ctacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5461 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5521 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5581 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5641 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5701 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgtttcc caatgatgag
     5761 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     5821 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5881 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5941 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6001 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     6061 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     6121 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     6181 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     6241 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     6301 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6361 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6421 gtcagaccaa gtttactcat atatacttta gattgattta ccccggttga taatcagaaa
     6481 agccccaaaa acaggaagat tgtataagca aatatttaaa ttgtaaacgt taatattttg
     6541 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     6601 ggcaaaatcc cttataaatc aaaagaatag cccgagatag ggttgagtgt tgttccagtt
     6661 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     6721 tatcagggcg atggcccact acgtgaacca tcacccaaat caagtttttt ggggtcgagg
     6781 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     6841 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     6901 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     6961 ctacagggcg cgtaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7021 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7081 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7141 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7201 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     7261 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7321 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     7381 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     7441 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     7501 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     7561 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     7621 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     7681 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     7741 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     7801 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcta tggtgcactc
     7861 tcagtacaat ctgctctgat gccgcatagt taagccagt
//
