LOCUS       sequence-354811-        8586 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3-component ShHELIX containing a Cas12k-TniQ fusion. New 
            sgRNA spacer sequences can be added using Golden Gate assembly (via 
            SapI sites)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8586)
  AUTHORS   Tou CJ, Orr B, Kleinstiver BP
  TITLE     Precise cut-and-paste DNA insertion using engineered type V-K 
            CRISPR-associated transposases.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01574-x.
  PUBMED    36593413
REFERENCE   2  (bases 1 to 8586)
  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. 2023 Jan 2. doi: 10.1038/s41587-022-01574-x."
COMMENT     Sequence Label: pShHELIX(Cas12k-TniQ)-sgRNA_entry (CJT111)
FEATURES             Location/Qualifiers
     source          1..8586
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      2686..2757
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        2771..2805
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     terminator      3060..3087
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3114..3218
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3219..3287,3288..4079)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3437..3456)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4250..4838
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4739..4758
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(4938..4943,4944..4961,4962..4968)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             4969..4980
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 ctgtggtatt cactgttggc acgacaacac ctttatcgcg ccaatactgt agctgacgac
       61 gcgaacaatt agtgatttga gccgcttctg tgcttgtgta gaattttcct tccatagaaa
      121 tcatcctaca gaagtagttt tttaaattta cattttttaa aagcacatta tgtattcatg
      181 tgatattatt ttttcatgag tcaaataact attcaagctc gacttatttc ctttgaatca
      241 aaccgccaac aactctggaa gttgatggca gatttaaaca cgccgttaat taacgaactg
      301 ctttgccagt taggtcaaca ccccgacttc gagaagtggc aacaaaaggg taaactcccg
      361 tctaccgttg tgagccagtt atgtcaacct ctcaaaactg accctcgctt tgcaggtcag
      421 cccagccgtt tatatatgtc ggcaattcat attgtggact acatctacaa gtcctggctg
      481 gctatacaga aacggcttca acagcagcta gatggaaaga cgcgctggct agaaatgctc
      541 aatagcgatg ctgaattagt agaacttagt ggtgacactt tagaggctat tcgtgtcaaa
      601 gctgctgaaa ttttggcaat agctatgcca gcatctgagt cagatagcgc ttcacctaaa
      661 gggaaaaaag gtaaaaagga gaaaaaaccc tcatcttcta gccctaagcg tagtttatcc
      721 aagacattat ttgacgctta ccaagaaacg gaagatatca agagccgtag cgccatcagc
      781 tacctgttaa aaaatggctg caaacttact gacaaagaag aagattcaga aaaatttgct
      841 aaacgtcgtc gtcaagttga aatccaaatt caaaggctta ccgaaaagtt aataagtcgg
      901 atgcctaaag gtcgagattt gaccaatgct aaatggttgg agacactctt gactgctaca
      961 accactgttg ctgaagacaa cgcccaagcc aaacgctggc aggatattct gttaactcga
     1021 tcaagttctc tcccattccc ccttgttttt gaaaccaacg aggatatggt ttggtcaaag
     1081 aatcaaaagg gtaggctgtg tgttcacttc aatggcttaa gcgatttaat ttttgaggtg
     1141 tactgcggca atcgtcaact tcactggttt caacgcttcc tagaagacca acagactaaa
     1201 cgcaaaagca aaaatcagca ttctagcggc ttgttcacac tcagaaatgg tcatctagtt
     1261 tggcttgaag gtgagggtaa aggggaacct tggaatcttc accacttgac cctttactgc
     1321 tgtgttgaca atcgcttgtg gacagaggag ggaacagaaa tcgttcgcca agagaaagca
     1381 gatgaaatta ctaaattcat cacaaacatg aagaagaaaa gcgatctaag cgatacacag
     1441 caagctttga ttcaacgtaa acaatcaaca cttactcgaa taaacaattc ctttgagcgt
     1501 cctagccaac ccctttatca aggtcaatca cacattttgg ttggagtaag cctgggacta
     1561 gaaaaacctg ccacagtagc agtagtagat gcgatcgcca acaaagtctt ggcttaccgg
     1621 agtattaaac aattacttgg cgacaattac gaactgctaa atcgccagag acgacaacag
     1681 cagtacctat ctcacgaacg ccacaaagca caaaaaaact tctctcccaa tcaatttgga
     1741 gcatctgagt tagggcaaca tatagacaga ttattagcta aagcaattgt agcgttagcg
     1801 agaacctaca aagctggcag tattgtcttg cccaagttag gggatatgcg ggaggttgtc
     1861 caaagtgaaa ttcaagctat agcagaacaa aaatttcccg gttatattga aggtcagcaa
     1921 aaatatgcca aacagtaccg ggttaatgtt catcggtgga gctacggcag attaattcaa
     1981 agcattcaaa gtaaagcagc tcaaacagga attgtgattg aggagggaaa acaacctatt
     2041 cgaggtagtc cccacgacaa agcaaaggaa ttagcacttt ctgcttacaa tctccgccta
     2101 actaggcgaa gtagcggcag cgagactccc gggacctcag agtccgccac acccgaaagt
     2161 gggagcatag aagcaccaga tgttaaacct tggctattct tgattaaacc ctatgaaggg
     2221 gaaagcctga gccactttct tggcaggttc agacgtgcca accatttatc cgcaagtgga
     2281 ttgggtactt tggcaggaat tggtgctata gtggcacgtt gggaaagatt tcattttaat
     2341 cctcgcccta gtcagcaaga attggaagcg atcgcatctg tagtagaagt ggatgctcaa
     2401 aggttagccc agatgttacc gcctgctgga gtgggaatgc agcatgagcc aattcgcttg
     2461 tgtggggctt gttatgccga gtcgccttgt caccgaattg aatggcagta caagtcggtg
     2521 tggaagtgcg atcgccatca actcaagatt ttagcaaagt gtccaaactg tcaagcacct
     2581 tttaaaatgc ctgcgctgtg ggaggatggg tgctgtcaca gatgtaggat gccgtttgca
     2641 gaaatggcaa agctacagaa ggtttgagta aggatctcca ggcatcaaat aaaacgaaag
     2701 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac
     2761 tagagagacc ctgacagcta gctcagtcct aggtataatg ctagcatatt aatagcgccg
     2821 caattcatgc tgcttgcagc ctctgaattt tgttaaatga gggttagttt gactgtataa
     2881 atacagtctt gctttctgac cctggtagct gctcaccctg atgctgctgt caatagacag
     2941 gataggtgcg ctcccagcaa taagggcgcg gatgtactgc tgtagtggct actgaatcac
     3001 ccccgatcaa gggggaaccc taaatgggtt gaaaggaaga gcgagatctc gagctcttcg
     3061 gctcaccttc gggtgggcct ttctgcgcag gtggcacttt tcggggaaat gtgcgcggaa
     3121 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3181 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3241 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3301 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3361 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3421 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3481 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3541 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3601 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3661 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3721 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3781 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3841 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3901 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3961 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4021 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4081 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4141 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4201 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4261 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4321 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4381 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4441 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4501 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4561 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4621 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4681 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4741 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4801 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4861 tacggttcct ggccttttgc tggccttttg ctcaacgcca agctccaaaa cgatctcaag
     4921 aagatcatct tattaatttt acactttatg cttccggctc gtatgttgaa agaggagaaa
     4981 ggatctatgg gcagcgattt aacatacgca tatctagtag gcctgtttga gggtgatggt
     5041 tatttttcca ttacgaaaaa aggtaaatac cttacctatg agctggggat cgagctgagc
     5101 atcaaggacg tgcagctgat ctataaaatc aagaagattc tgggcatcgg cattgtaagc
     5161 ttccgtaaac gcaatgaaat tgagatggtg gcgctgcgca ttcgcgataa gaaccacctg
     5221 aaaagcaaaa ttttgccaat cttcgagaaa tacccgatgt tttctaacaa acaatacgac
     5281 tatctgcgtt tccgcaacgc tttgctgagc ggtattatct ccctggaaga tctgccggat
     5341 tacacgcgtt ctgatgaacc gctgaacagc attgagagca tcattaacac cagctatttc
     5401 agtgcgtggc tggttggttt tatcgaggct gaaggctgct tttccgtgta caagttaaac
     5461 aaggatgacg actacttgat cgcgagcttc gacattgcgc aacgtgacgg cgacatcctc
     5521 atctcggcca ttcgtaagta cttgtcattc accactaaag tttacttaga taaaaccaat
     5581 tgtagcaagt tgaaggtcac ctctgttcgt tccgtggaaa acattatcaa attcctgcaa
     5641 aacgcaccgg ttaaattgtt gggtaatatg aaactgcagt ataagctgtg gctgaagcag
     5701 ctgagaaaga tttctcgtta cagcgaaaag atcaagatcc cgagcaatta tagcggcagc
     5761 gagactcccg ggacctcaga gtccgccaca cccgaaagtg ggagcaacag tcagcaaaat
     5821 cctgatttag ctgttcatcc cttggcaatt cctatggaag gcttactagg agaaagtgct
     5881 acaactcttg agaagaatgt aattgccaca caactctcag aggaagccca agtaaagcta
     5941 gaggtaatcc aaagtttact ggaaccctgc gatcgcacaa cttatgggca aaagttgcgg
     6001 gaagcagcag agaaactaaa tgtatcgttg cgaacggtac aaaggttggt gaaaaactgg
     6061 gaacaagatg gcttagtcgg actcactcaa acaagtaggg ctgataaagg aaaacaccgc
     6121 attggtgagt tttgggaaaa cttcattacc aaaacctaca aggagggtaa caagggaagt
     6181 aaacgtatga cccctaaaca agttgctctc agagtcgagg ctaaagcccg tgaattaaaa
     6241 gactctaagc cgcccaatta caaaaccgtg ttacgggtat tagcacccat tttggaaaag
     6301 caacaaaaag ccaagagtat ccgcagtcct ggttggagag gaactacgct ttcggttaaa
     6361 acccgtgaag gaaaagattt atcggttgat tacagtaacc atgtttggca atgtgaccat
     6421 acccgcgtgg atgtgttgct ggtagatcaa catggtgaaa ttttaagtcg tccctggcta
     6481 acaacagtaa ttgatactta ctctcgttgc attatgggta tcaacttggg ctttgatgca
     6541 cccagttctg gggtagtagc attagcgtta cgccatgcaa ttctaccaaa gcgttacggt
     6601 tccgagtaca aactgcattg tgagtgggga acctatggaa aaccagaaca tttttatact
     6661 gatggcggta aagactttcg ctctaaccac ttgagtcaga ttggggcgca attgggattt
     6721 gtctgtcatt tacgcgatcg cccttctgaa ggtggagtag tagaacgtcc cttcaaaaca
     6781 ttaaatgacc aactattttc aacgcttcct gggtacaccg gatctaatgt gcaggaacgc
     6841 ccagaagatg cagagaagga cgcaagactt actttgcgag aactagaaca gttacttgtg
     6901 cgttacatcg tagatcgtta caaccaaagt attgatgcgc ggatgggcga ccaaacgcgc
     6961 tttgagcgtt gggaagcagg attgcctaca gtgccagtac caataccaga acgagatttg
     7021 gatatttgtt taatgaagca gtcacggcgc actgtgcaaa gaggtggttg tttgcagttt
     7081 cagaatttaa tgtatcgggg ggaatatttg gcaggttatg ccggagaaac tgtcaactta
     7141 aggtttgacc ccagagacat tacaacaatt ttggtttatc gccaggaaaa caatcaggaa
     7201 gtatttctga ctcgcgctca cgctcaaggt ttggagacag agcaactggc attagatgag
     7261 gctgaggcag caagtcgcag actccgtacc gcagggaaaa ctatcagtaa ccaatcatta
     7321 ttgcaagaag ttgttgaccg cgatgctctt gtcgctacca agaaaagccg taaggagcgt
     7381 caaaaattgg aacagactgt tttgcgatct gctgctgttg atgaaagtaa tagagaatcc
     7441 ttgccttctc aaatagttga accagatgaa gtggaatcta cagaaacggt tcactctcaa
     7501 tacgaagaca ttgaggtgtg ggactatgaa caacttcgtg aagaatatgg gttttaaaca
     7561 atgacagaag ctcaggcgat cgccaagcag ttgggtgggg taaaaccgga tgatgagtgg
     7621 ttacaagctg aaattgctcg tctcaagggt aagagcattg tgcctttaca gcaggtaaaa
     7681 actctccatg attggttaga tggcaagcgc aaggcaagaa aatcttgccg agtagttggg
     7741 gaatcgagaa ctggcaagac agttgcttgt gatgcctaca gatacaggca caaacctcag
     7801 caggaagctg gacgacctcc aactgtgcct gtcgtttata ttcgacctca ccaaaaatgt
     7861 ggccccaagg atttgtttaa aaagattact gagtacctca agtatcgggt aacaaaaggg
     7921 actgtatctg attttcgaga taggacgata gaagtactca agggttgtgg cgtagagatg
     7981 ctaattattg atgaagctga ccgtctcaag cctgaaactt ttgctgatgt gcgagatatt
     8041 gccgaagatt taggaattgc tgtggtactg gtaggaacag accgtttgga tgcggtaatt
     8101 aagcgggatg agcaggttct cgaacgcttt cgggcgcatc ttcgctttgg taaattgtcg
     8161 ggagaggatt ttaagaacac cgtagaaatg tgggaacaaa tggttttgaa actgccagta
     8221 tcttctaatc taaagagcaa ggagatgcta cggattctca cgtcagcaac tgaaggctac
     8281 attggtcgcc ttgatgagat tcttagggaa gctgcaattc gttccttatc aagaggattg
     8341 aagaagattg acaaggctgt tttacaggaa gtagctaagg agtacaaatg actgcagtga
     8401 taaaaccaga aaaaggtgtg aaattaacta agtccctgaa ttgatctggt tgtccaaaaa
     8461 atttgtgcga tcgcatggca agattattcc tactattgat gtggcgtgag tcggataact
     8521 tgctctaatg ggatcccggt taataccaat agatcagata tgagtaataa acattacgac
     8581 ctttac
//
