LOCUS       sequence-340458-        7483 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible expression of Transposon TnY fused to Mxe Intein and 
            Chitin-binding domain..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7483)
  AUTHORS   Liscovitch-Brauer N, Montalbano A, Deng J, Mendez-Mancilla A, 
            Wessels HH, Moss NG, Kung CY, Sookdeo A, Guo X, Geller E, Jaini S, 
            Smibert P, Sanjana NE
  TITLE     Profiling the genetic determinants of chromatin accessibility with 
            scalable single-cell CRISPR screens.
  JOURNAL   Nat Biotechnol. 2021 Apr 29. pii: 10.1038/s41587-021-00902-x. doi: 
            10.1038/s41587-021-00902-x.
  PUBMED    33927415
REFERENCE   2  (bases 1 to 7483)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2021 Apr 29. pii: 10.1038/s41587-021-00902-x. doi: 
            10.1038/s41587-021-00902-x."
COMMENT     Sequence Label: pTXB1-TnY
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1466..2059
                     /codon_start=1
                     /gene="Mycobacterium xenopi gyrA"
                     /product="modified GyrA intein (Southworth et al., 1999)"
                     /label=Mxe GyrA intein
                     /note="N-terminal cleavage of this intein can be induced by
                     thiols."
                     /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
                     LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
                     IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
                     AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
     CDS             2090..2245
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     primer_bind     complement(2310..2328)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2324..2371
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2431..2535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2536..2604,2605..3396)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2754..2773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3438..3951)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3633..3654)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3845..3864
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4062..4650
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4551..4570
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4804..4821
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(4843..4862)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5003..5025
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5019..5210)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5767..6849)
                     /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"
     primer_bind     6811..6830
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6850..6927)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      7080..7166
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7263..7349
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gactcacagc gatgcgaaac tgtgggctca
      121 ggagcaattc ggtcaggccc aactgaaaga tccgcgccgc acccagcgcc tgatttctct
      181 ggcgaccagc attgctaacc agccgggtgt tagcgttgcg aaactgccgt tttctaaagc
      241 cgatcaggag ggcgcgtacc gtttcattcg taacgataac atcgacgcga aagacatcgc
      301 tgaagcaggc tttcagtcca ccgtatcccg cgctaacgaa cacaaagagc tgctggcgct
      361 ggaagacact acgaccctgt ctttcccgca tcgttccatc aaagaagaac tgggccatac
      421 gaaccagggt gatcgcaccc gcgccctgca cgttcactct accctgctgt tcgcgccgca
      481 gaaccagact atcgtgggtc tgatcgagca gcagcgttgg tctcgtgata ttactaaacg
      541 cggtcagaaa catcagcacg ctacccgtcc ttataaagaa aaagaatcct ataaatggga
      601 gcaggcttcc cgtcgtgttg tggagcgcct gggtgataaa atgctggatg tcatttctgt
      661 ttgcgaccgc gaggcagatc tgtttgaata cctgacctac aaacgtcaac accagcagcg
      721 tttcgttgtt cgtagcatgc agtctcgctg tctggaagaa cacgctcaga aactgtatga
      781 ctacgcacag gcgctgccat ctgtaaaaac gaaggcactg accatccctc aaaaaggtgg
      841 ccgtaaagca cgtgacgtta aactggacgt taaatacggc caggttactc tgaaagcgcc
      901 ggccaacaaa aaggagcacg caggcattcc ggtttactac gtgggctgcc tggaacaggg
      961 tacttccaaa gataaactgg cgtggcacct gctgacctct gaacctatta acaacgtcga
     1021 ggatgccatg cgtatcatcg gctactacga acgtcgttgg ctgatcgagg attttcacaa
     1081 agtatggaaa tccgaaggta ctgacgtaga atccctgcgt ctgcagagca aagacaacct
     1141 ggaacgtctg tccgttatct acgcgtttgt tgctacccgc ctgctggcac tgcgttttat
     1201 caaggaagtt gatgaactga ccaaagaaag ctgtgaaaaa gttctgggcc agaaagcgtg
     1261 gaaactgctg tggctgaagc tggaatctaa aaccctgccg aaagaggtac cggacatggg
     1321 ttgggcttat aaaaacctgg ctaaactggg tggctggaag gacactaagc gtaccggtcg
     1381 cgcttctatc aaagttctgt gggagggttg gttcaaactg cagaccatcc tggagggcta
     1441 tgaactggcg atgtccctgg accactgcat cacgggagat gcactagttg ccctacccga
     1501 gggcgagtcg gtacgcatcg ccgacatcgt gccgggtgcg cggcccaaca gtgacaacgc
     1561 catcgacctg aaagtccttg accggcatgg caatcccgtg ctcgccgacc ggctgttcca
     1621 ctccggcgag catccggtgt acacggtgcg tacggtcgaa ggtctgcgtg tgacgggcac
     1681 cgcgaaccac ccgttgttgt gtttggtcga cgtcgccggg gtgccgaccc tgctgtggaa
     1741 gctgatcgac gaaatcaagc cgggcgatta cgcggtgatt caacgcagcg cattcagcgt
     1801 cgactgtgca ggttttgccc gcgggaaacc cgaatttgcg cccacaacct acacagtcgg
     1861 cgtccctgga ctggtgcgtt tcttggaagc acaccaccga gacccggacg cccaagctat
     1921 cgccgacgag ctgaccgacg ggcggttcta ctacgcgaaa gtcgccagtg tcaccgacgc
     1981 cggcgtgcag ccggtgtata gccttcgtgt cgacacggca gaccacgcgt ttatcacgaa
     2041 cgggttcgtc agccacgcta ctggcctcac cggtctgaac tcaggcctca cgacaaatcc
     2101 tggtgtatcc gcttggcagg tcaacacagc ttatactgcg ggacaattgg tcacatataa
     2161 cggcaagacg tataaatgtt tgcagcccca cacctccttg gcaggatggg aaccatccaa
     2221 cgttcctgcc ttgtggcagc ttcaatgact gcaggaaggg gatccggctg ctaacaaagc
     2281 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     2341 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggataact
     2401 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     2461 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     2521 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     2581 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     2641 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     2701 gagagttttc gccccgaaga acgtttccca atgatgagca cttttaaagt tctgctatgt
     2761 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     2821 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     2881 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2941 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3001 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3061 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3121 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3181 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3241 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     3301 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     3361 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     3421 atactttaga ttgatttacc ccggttgata atcagaaaag ccccaaaaac aggaagattg
     3481 tataagcaaa tatttaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt
     3541 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3601 aagaatagcc cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3661 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3721 gtgaaccatc acccaaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3781 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3841 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3901 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg taaaaggatc
     3961 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4021 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4081 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4141 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4201 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4261 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4321 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4381 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4441 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4501 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4561 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4621 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4681 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4741 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4801 cgcagcgagt cagtgagcga ggaagctatg gtgcactctc agtacaatct gctctgatgc
     4861 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     4921 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4981 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5041 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgcag cgattcacag
     5101 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     5161 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     5221 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     5281 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     5341 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagccgaac
     5401 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     5461 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     5521 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     5581 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
     5641 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     5701 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     5761 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5821 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     5881 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5941 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     6001 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     6061 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     6121 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     6181 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     6241 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     6301 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     6361 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     6421 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     6481 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     6541 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     6601 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     6661 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     6721 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     6781 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     6841 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     6901 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     6961 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     7021 gtgcatgccg gcatgccgcc ctttcgtctt caagaattaa ttcccaattc cccaggcatc
     7081 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     7141 tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac
     7201 ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggaattaa ttccccaggc
     7261 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     7321 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     7381 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggaat tggggatcgg
     7441 aattaattcc cggtttaaac cggggatctc gatcccgcga aat
//
