LOCUS       sequence-354814-        9120 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3-component ShHELIX containing a Cas12k-TniQ-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 9120)
  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 9120)
  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-TniQ)-sgRNA_entry (CJT112)
FEATURES             Location/Qualifiers
     source          1..9120
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      80..151
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        165..199
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     terminator      454..481
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        508..612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(613..681,682..1473)
                     /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(831..850)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1644..2232
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2133..2152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(2332..2337,2338..2355,2356..2362)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             2363..2374
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 tgggtgctgt cacagatgta ggatgccgtt tgcagaaatg gcaaagctac agaaggtttg
       61 agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      121 ttttatctgt tgtttgtcgg tgaacgctct ctactagaga gaccctgaca gctagctcag
      181 tcctaggtat aatgctagca tattaatagc gccgcaattc atgctgcttg cagcctctga
      241 attttgttaa atgagggtta gtttgactgt ataaatacag tcttgctttc tgaccctggt
      301 agctgctcac cctgatgctg ctgtcaatag acaggatagg tgcgctccca gcaataaggg
      361 cgcggatgta ctgctgtagt ggctactgaa tcacccccga tcaaggggga accctaaatg
      421 ggttgaaagg aagagcgaga tctcgagctc ttcggctcac cttcgggtgg gcctttctgc
      481 gcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1141 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1321 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1621 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1681 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1741 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1801 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1861 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1921 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1981 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2041 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2101 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2161 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2221 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2281 tttgctcaac gccaagctcc aaaacgatct caagaagatc atcttattaa ttttacactt
     2341 tatgcttccg gctcgtatgt tgaaagagga gaaaggatct atgggcagcg atttaacata
     2401 cgcatatcta gtaggcctgt ttgagggtga tggttatttt tccattacga aaaaaggtaa
     2461 ataccttacc tatgagctgg ggatcgagct gagcatcaag gacgtgcagc tgatctataa
     2521 aatcaagaag attctgggca tcggcattgt aagcttccgt aaacgcaatg aaattgagat
     2581 ggtggcgctg cgcattcgcg ataagaacca cctgaaaagc aaaattttgc caatcttcga
     2641 gaaatacccg atgttttcta acaaacaata cgactatctg cgtttccgca acgctttgct
     2701 gagcggtatt atctccctgg aagatctgcc ggattacacg cgttctgatg aaccgctgaa
     2761 cagcattgag agcatcatta acaccagcta tttcagtgcg tggctggttg gttttatcga
     2821 ggctgaaggc tgcttttccg tgtacaagtt aaacaaggat gacgactact tgatcgcgag
     2881 cttcgacatt gcgcaacgtg acggcgacat cctcatctcg gccattcgta agtacttgtc
     2941 attcaccact aaagtttact tagataaaac caattgtagc aagttgaagg tcacctctgt
     3001 tcgttccgtg gaaaacatta tcaaattcct gcaaaacgca ccggttaaat tgttgggtaa
     3061 tatgaaactg cagtataagc tgtggctgaa gcagctgaga aagatttctc gttacagcga
     3121 aaagatcaag atcccgagca attatagcgg cagcgagact cccgggacct cagagtccgc
     3181 cacacccgaa agtgggagca acagtcagca aaatcctgat ttagctgttc atcccttggc
     3241 aattcctatg gaaggcttac taggagaaag tgctacaact cttgagaaga atgtaattgc
     3301 cacacaactc tcagaggaag cccaagtaaa gctagaggta atccaaagtt tactggaacc
     3361 ctgcgatcgc acaacttatg ggcaaaagtt gcgggaagca gcagagaaac taaatgtatc
     3421 gttgcgaacg gtacaaaggt tggtgaaaaa ctgggaacaa gatggcttag tcggactcac
     3481 tcaaacaagt agggctgata aaggaaaaca ccgcattggt gagttttggg aaaacttcat
     3541 taccaaaacc tacaaggagg gtaacaaggg aagtaaacgt atgaccccta aacaagttgc
     3601 tctcagagtc gaggctaaag cccgtgaatt aaaagactct aagccgccca attacaaaac
     3661 cgtgttacgg gtattagcac ccattttgga aaagcaacaa aaagccaaga gtatccgcag
     3721 tcctggttgg agaggaacta cgctttcggt taaaacccgt gaaggaaaag atttatcggt
     3781 tgattacagt aaccatgttt ggcaatgtga ccatacccgc gtggatgtgt tgctggtaga
     3841 tcaacatggt gaaattttaa gtcgtccctg gctaacaaca gtaattgata cttactctcg
     3901 ttgcattatg ggtatcaact tgggctttga tgcacccagt tctggggtag tagcattagc
     3961 gttacgccat gcaattctac caaagcgtta cggttccgag tacaaactgc attgtgagtg
     4021 gggaacctat ggaaaaccag aacattttta tactgatggc ggtaaagact ttcgctctaa
     4081 ccacttgagt cagattgggg cgcaattggg atttgtctgt catttacgcg atcgcccttc
     4141 tgaaggtgga gtagtagaac gtcccttcaa aacattaaat gaccaactat tttcaacgct
     4201 tcctgggtac accggatcta atgtgcagga acgcccagaa gatgcagaga aggacgcaag
     4261 acttactttg cgagaactag aacagttact tgtgcgttac atcgtagatc gttacaacca
     4321 aagtattgat gcgcggatgg gcgaccaaac gcgctttgag cgttgggaag caggattgcc
     4381 tacagtgcca gtaccaatac cagaacgaga tttggatatt tgtttaatga agcagtcacg
     4441 gcgcactgtg caaagaggtg gttgtttgca gtttcagaat ttaatgtatc ggggggaata
     4501 tttggcaggt tatgccggag aaactgtcaa cttaaggttt gaccccagag acattacaac
     4561 aattttggtt tatcgccagg aaaacaatca ggaagtattt ctgactcgcg ctcacgctca
     4621 aggtttggag acagagcaac tggcattaga tgaggctgag gcagcaagtc gcagactccg
     4681 taccgcaggg aaaactatca gtaaccaatc attattgcaa gaagttgttg accgcgatgc
     4741 tcttgtcgct accaagaaaa gccgtaagga gcgtcaaaaa ttggaacaga ctgttttgcg
     4801 atctgctgct gttgatgaaa gtaatagaga atccttgcct tctcaaatag ttgaaccaga
     4861 tgaagtggaa tctacagaaa cggttcactc tcaatacgaa gacattgagg tgtgggacta
     4921 tgaacaactt cgtgaagaat atgggtttta aacaatgaca gaagctcagg cgatcgccaa
     4981 gcagttgggt ggggtaaaac cggatgatga gtggttacaa gctgaaattg ctcgtctcaa
     5041 gggtaagagc attgtgcctt tacagcaggt aaaaactctc catgattggt tagatggcaa
     5101 gcgcaaggca agaaaatctt gccgagtagt tggggaatcg agaactggca agacagttgc
     5161 ttgtgatgcc tacagataca ggcacaaacc tcagcaggaa gctggacgac ctccaactgt
     5221 gcctgtcgtt tatattcgac ctcaccaaaa atgtggcccc aaggatttgt ttaaaaagat
     5281 tactgagtac ctcaagtatc gggtaacaaa agggactgta tctgattttc gagataggac
     5341 gatagaagta ctcaagggtt gtggcgtaga gatgctaatt attgatgaag ctgaccgtct
     5401 caagcctgaa acttttgctg atgtgcgaga tattgccgaa gatttaggaa ttgctgtggt
     5461 actggtagga acagaccgtt tggatgcggt aattaagcgg gatgagcagg ttctcgaacg
     5521 ctttcgggcg catcttcgct ttggtaaatt gtcgggagag gattttaaga acaccgtaga
     5581 aatgtgggaa caaatggttt tgaaactgcc agtatcttct aatctaaaga gcaaggagat
     5641 gctacggatt ctcacgtcag caactgaagg ctacattggt cgccttgatg agattcttag
     5701 ggaagctgca attcgttcct tatcaagagg attgaagaag attgacaagg ctgttttaca
     5761 ggaagtagct aaggagtaca aatgactgca gtgataaaac cagaaaaagg tgtgaaatta
     5821 actaagtccc tgaattgatc tggttgtcca aaaaatttgt gcgatcgcat ggcaagatta
     5881 ttcctactat tgatgtggcg tgagtcggat aacttgctct aatgggatcc cggttaatac
     5941 caatagatca gatatgagta ataaacatta cgacctttac ctgtggtatt cactgttggc
     6001 acgacaacac ctttatcgcg ccaatactgt agctgacgac gcgaacaatt agtgatttga
     6061 gccgcttctg tgcttgtgta gaattttcct tccatagaaa tcatcctaca gaagtagttt
     6121 tttaaattta cattttttaa aagcacatta tgtattcatg tgatattatt ttttcatgag
     6181 tcaaataact attcaagctc gacttatttc ctttgaatca aaccgccaac aactctggaa
     6241 gttgatggca gatttaaaca cgccgttaat taacgaactg ctttgccagt taggtcaaca
     6301 ccccgacttc gagaagtggc aacaaaaggg taaactcccg tctaccgttg tgagccagtt
     6361 atgtcaacct ctcaaaactg accctcgctt tgcaggtcag cccagccgtt tatatatgtc
     6421 ggcaattcat attgtggact acatctacaa gtcctggctg gctatacaga aacggcttca
     6481 acagcagcta gatggaaaga cgcgctggct agaaatgctc aatagcgatg ctgaattagt
     6541 agaacttagt ggtgacactt tagaggctat tcgtgtcaaa gctgctgaaa ttttggcaat
     6601 agctatgcca gcatctgagt cagatagcgc ttcacctaaa gggaaaaaag gtaaaaagga
     6661 gaaaaaaccc tcatcttcta gccctaagcg tagtttatcc aagacattat ttgacgctta
     6721 ccaagaaacg gaagatatca agagccgtag cgccatcagc tacctgttaa aaaatggctg
     6781 caaacttact gacaaagaag aagattcaga aaaatttgct aaacgtcgtc gtcaagttga
     6841 aatccaaatt caaaggctta ccgaaaagtt aataagtcgg atgcctaaag gtcgagattt
     6901 gaccaatgct aaatggttgg agacactctt gactgctaca accactgttg ctgaagacaa
     6961 cgcccaagcc aaacgctggc aggatattct gttaactcga tcaagttctc tcccattccc
     7021 ccttgttttt gaaaccaacg aggatatggt ttggtcaaag aatcaaaagg gtaggctgtg
     7081 tgttcacttc aatggcttaa gcgatttaat ttttgaggtg tactgcggca atcgtcaact
     7141 tcactggttt caacgcttcc tagaagacca acagactaaa cgcaaaagca aaaatcagca
     7201 ttctagcggc ttgttcacac tcagaaatgg tcatctagtt tggcttgaag gtgagggtaa
     7261 aggggaacct tggaatcttc accacttgac cctttactgc tgtgttgaca atcgcttgtg
     7321 gacagaggag ggaacagaaa tcgttcgcca agagaaagca gatgaaatta ctaaattcat
     7381 cacaaacatg aagaagaaaa gcgatctaag cgatacacag caagctttga ttcaacgtaa
     7441 acaatcaaca cttactcgaa taaacaattc ctttgagcgt cctagccaac ccctttatca
     7501 aggtcaatca cacattttgg ttggagtaag cctgggacta gaaaaacctg ccacagtagc
     7561 agtagtagat gcgatcgcca acaaagtctt ggcttaccgg agtattaaac aattacttgg
     7621 cgacaattac gaactgctaa atcgccagag acgacaacag cagtacctat ctcacgaacg
     7681 ccacaaagca caaaaaaact tctctcccaa tcaatttgga gcatctgagt tagggcaaca
     7741 tatagacaga ttattagcta aagcaattgt agcgttagcg agaacctaca aagctggcag
     7801 tattgtcttg cccaagttag gggatatgcg ggaggttgtc caaagtgaaa ttcaagctat
     7861 agcagaacaa aaatttcccg gttatattga aggtcagcaa aaatatgcca aacagtaccg
     7921 ggttaatgtt catcggtgga gctacggcag attaattcaa agcattcaaa gtaaagcagc
     7981 tcaaacagga attgtgattg aggagggaaa acaacctatt cgaggtagtc cccacgacaa
     8041 agcaaaggaa ttagcacttt ctgcttacaa tctccgccta actaggcgaa gtagcggcag
     8101 cgagactccc gggacctcag agtccgccac acccgaaagt gggagcatag aagcaccaga
     8161 tgttaaacct tggctattct tgattaaacc ctatgaaggg gaaagcctga gccactttct
     8221 tggcaggttc agacgtgcca accatttatc cgcaagtgga ttgggtactt tggcaggaat
     8281 tggtgctata gtggcacgtt gggaaagatt tcattttaat cctcgcccta gtcagcaaga
     8341 attggaagcg atcgcatctg tagtagaagt ggatgctcaa aggttagccc agatgttacc
     8401 gcctgctgga gtgggaatgc agcatgagcc aattcgcttg tgtggggctt gttatgccga
     8461 gtcgccttgt caccgaattg aatggcagta caagtcggtg tggaagtgcg atcgccatca
     8521 actcaagatt ttagcaaagt gtccaaactg tcaagcacct tttaaaatgc ctgcgctgtg
     8581 ggaggatggg tgctgtcaca gatgtaggat gccgtttgca gaaatggcaa agctacagaa
     8641 ggttggcggt ggctctggag gtgggtccgg cggtggctct atagaagcac cagatgttaa
     8701 accttggcta ttcttgatta aaccctatga aggggaaagc ctgagccact ttcttggcag
     8761 gttcagacgt gccaaccatt tatccgcaag tggattgggt actttggcag gaattggtgc
     8821 tatagtggca cgttgggaaa gatttcattt taatcctcgc cctagtcagc aagaattgga
     8881 agcgatcgca tctgtagtag aagtggatgc tcaaaggtta gcccagatgt taccgcctgc
     8941 tggagtggga atgcagcatg agccaattcg cttgtgtggg gcttgttatg ccgagtcgcc
     9001 ttgtcaccga attgaatggc agtacaagtc ggtgtggaag tgcgatcgcc atcaactcaa
     9061 gattttagca aagtgtccaa actgtcaagc accttttaaa atgcctgcgc tgtgggagga
//
