LOCUS       sequence-245554-        8156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ShCAST and a crRNA targeting PSP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8156)
  AUTHORS   Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin
            EV, Zhang F
  TITLE     RNA-guided DNA insertion with CRISPR-associated transposases.
  JOURNAL   Science. 2019 Jul 5;365(6448):48-53. doi: 10.1126/science.aax9181. 
            Epub 2019 Jun 6.
  PUBMED    31171706
REFERENCE   2  (bases 1 to 8156)
  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"; doi: 
            "10.1126/science.aax9181"; journalName: "Science"; date: "2019-07-5-
            5"; volume: "365"; issue: "6448"; pages: "48-53"
COMMENT     Sequence Label: pHelper_ShCAST
FEATURES             Location/Qualifiers
     source          1..8156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(82..87,88..105,106..112)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             113..124
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        5975..6009
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(6149..6168)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6268..6290
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6328..6346)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6414..6518
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6519..6587,6588..7379)
                     /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(6737..6756)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7550..8138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8039..8058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaac gccaagctcc aaaacgatct
       61 caagaagatc atcttattaa ttttacactt tatgcttccg gctcgtatgt tgaaagagga
      121 gaaaggatct atgaacagtc agcaaaatcc tgatttagct gttcatccct tggcaattcc
      181 tatggaaggc ttactaggag aaagtgctac aactcttgag aagaatgtaa ttgccacaca
      241 actctcagag gaagcccaag taaagctaga ggtaatccaa agtttactgg aaccctgcga
      301 tcgcacaact tatgggcaaa agttgcggga agcagcagag aaactaaatg tatcgttgcg
      361 aacggtacaa aggttggtga aaaactggga acaagatggc ttagtcggac tcactcaaac
      421 aagtagggct gataaaggaa aacaccgcat tggtgagttt tgggaaaact tcattaccaa
      481 aacctacaag gagggtaaca agggaagtaa acgtatgacc cctaaacaag ttgctctcag
      541 agtcgaggct aaagcccgtg aattaaaaga ctctaagccg cccaattaca aaaccgtgtt
      601 acgggtatta gcacccattt tggaaaagca acaaaaagcc aagagtatcc gcagtcctgg
      661 ttggagagga actacgcttt cggttaaaac ccgtgaagga aaagatttat cggttgatta
      721 cagtaaccat gtttggcaat gtgaccatac ccgcgtggat gtgttgctgg tagatcaaca
      781 tggtgaaatt ttaagtcgtc cctggctaac aacagtaatt gatacttact ctcgttgcat
      841 tatgggtatc aacttgggct ttgatgcacc cagttctggg gtagtagcat tagcgttacg
      901 ccatgcaatt ctaccaaagc gttacggttc cgagtacaaa ctgcattgtg agtggggaac
      961 ctatggaaaa ccagaacatt tttatactga tggcggtaaa gactttcgct ctaaccactt
     1021 gagtcagatt ggggcgcaat tgggatttgt ctgtcattta cgcgatcgcc cttctgaagg
     1081 tggagtagta gaacgtccct tcaaaacatt aaatgaccaa ctattttcaa cgcttcctgg
     1141 gtacaccgga tctaatgtgc aggaacgccc agaagatgca gagaaggacg caagacttac
     1201 tttgcgagaa ctagaacagt tacttgtgcg ttacatcgta gatcgttaca accaaagtat
     1261 tgatgcgcgg atgggcgacc aaacgcgctt tgagcgttgg gaagcaggat tgcctacagt
     1321 gccagtacca ataccagaac gagatttgga tatttgttta atgaagcagt cacggcgcac
     1381 tgtgcaaaga ggtggttgtt tgcagtttca gaatttaatg tatcgggggg aatatttggc
     1441 aggttatgcc ggagaaactg tcaacttaag gtttgacccc agagacatta caacaatttt
     1501 ggtttatcgc caggaaaaca atcaggaagt atttctgact cgcgctcacg ctcaaggttt
     1561 ggagacagag caactggcat tagatgaggc tgaggcagca agtcgcagac tccgtaccgc
     1621 agggaaaact atcagtaacc aatcattatt gcaagaagtt gttgaccgcg atgctcttgt
     1681 cgctaccaag aaaagccgta aggagcgtca aaaattggaa cagactgttt tgcgatctgc
     1741 tgctgttgat gaaagtaata gagaatcctt gccttctcaa atagttgaac cagatgaagt
     1801 ggaatctaca gaaacggttc actctcaata cgaagacatt gaggtgtggg actatgaaca
     1861 acttcgtgaa gaatatgggt tttaaacaat gacagaagct caggcgatcg ccaagcagtt
     1921 gggtggggta aaaccggatg atgagtggtt acaagctgaa attgctcgtc tcaagggtaa
     1981 gagcattgtg cctttacagc aggtaaaaac tctccatgat tggttagatg gcaagcgcaa
     2041 ggcaagaaaa tcttgccgag tagttgggga atcgagaact ggcaagacag ttgcttgtga
     2101 tgcctacaga tacaggcaca aacctcagca ggaagctgga cgacctccaa ctgtgcctgt
     2161 cgtttatatt cgacctcacc aaaaatgtgg ccccaaggat ttgtttaaaa agattactga
     2221 gtacctcaag tatcgggtaa caaaagggac tgtatctgat tttcgagata ggacgataga
     2281 agtactcaag ggttgtggcg tagagatgct aattattgat gaagctgacc gtctcaagcc
     2341 tgaaactttt gctgatgtgc gagatattgc cgaagattta ggaattgctg tggtactggt
     2401 aggaacagac cgtttggatg cggtaattaa gcgggatgag caggttctcg aacgctttcg
     2461 ggcgcatctt cgctttggta aattgtcggg agaggatttt aagaacaccg tagaaatgtg
     2521 ggaacaaatg gttttgaaac tgccagtatc ttctaatcta aagagcaagg agatgctacg
     2581 gattctcacg tcagcaactg aaggctacat tggtcgcctt gatgagattc ttagggaagc
     2641 tgcaattcgt tccttatcaa gaggattgaa gaagattgac aaggctgttt tacaggaagt
     2701 agctaaggag tacaaatgat agaagcacca gatgttaaac cttggctatt cttgattaaa
     2761 ccctatgaag gggaaagcct gagccacttt cttggcaggt tcagacgtgc caaccattta
     2821 tccgcaagtg gattgggtac tttggcagga attggtgcta tagtggcacg ttgggaaaga
     2881 tttcatttta atcctcgccc tagtcagcaa gaattggaag cgatcgcatc tgtagtagaa
     2941 gtggatgctc aaaggttagc ccagatgtta ccgcctgctg gagtgggaat gcagcatgag
     3001 ccaattcgct tgtgtggggc ttgttatgcc gagtcgcctt gtcaccgaat tgaatggcag
     3061 tacaagtcgg tgtggaagtg cgatcgccat caactcaaga ttttagcaaa gtgtccaaac
     3121 tgtcaagcac cttttaaaat gcctgcgctg tgggaggatg ggtgctgtca cagatgtagg
     3181 atgccgtttg cagaaatggc aaagctacag aaggtttgac tgcagtgata aaaccagaaa
     3241 aaggtgtgaa attaactaag tccctgaatt gatctggttg tccaaaaaat ttgtgcgatc
     3301 gcatggcaag attattccta ctattgatgt ggcgtgagtc ggataacttg ctctaatggg
     3361 atcccggtta ataccaatag atcagatatg agtaataaac attacgacct ttacctgtgg
     3421 tattcactgt tggcacgaca acacctttat cgcgccaata ctgtagctga cgacgcgaac
     3481 aattagtgat ttgagccgct tctgtgcttg tgtagaattt tccttccata gaaatcatcc
     3541 tacagaagta gttttttaaa tttacatttt ttaaaagcac attatgtatt catgtgatat
     3601 tattttttta tgagtcaaat aactattcaa gctcgactta tttcctttga atcaaaccgc
     3661 caacaactct ggaagttgat ggcagattta aacacgccgt taattaacga actgctttgc
     3721 cagttaggtc aacaccccga cttcgagaag tggcaacaaa agggtaaact cccgtctacc
     3781 gttgtgagcc agttatgtca acctctcaaa actgaccctc gctttgcagg tcagcccagc
     3841 cgtttatata tgtcggcaat tcatattgtg gactacatct acaagtcctg gctggctata
     3901 cagaaacggc ttcaacagca gctagatgga aagacgcgct ggctagaaat gctcaatagc
     3961 gatgctgaat tagtagaact tagtggtgac actttagagg ctattcgtgt caaagctgct
     4021 gaaattttgg caatagctat gccagcatct gagtcagata gcgcttcacc taaagggaaa
     4081 aaaggtaaaa aggagaaaaa accctcatct tctagcccta agcgtagttt atccaagaca
     4141 ttatttgacg cttaccaaga aacggaagat atcaagagcc gtagcgccat cagctacctg
     4201 ttaaaaaatg gctgcaaact tactgacaaa gaagaagatt cagaaaaatt tgctaaacgt
     4261 cgtcgtcaag ttgaaatcca aattcaaagg cttaccgaaa agttaataag tcggatgcct
     4321 aaaggtcgag atttgaccaa tgctaaatgg ttggagacac tcttgactgc tacaaccact
     4381 gttgctgaag acaacgccca agccaaacgc tggcaggata ttctgttaac tcgatcaagt
     4441 tctctcccat tcccccttgt ttttgaaacc aacgaggata tggtttggtc aaagaatcaa
     4501 aagggtaggc tgtgtgttca cttcaatggc ttaagcgatt taatttttga ggtgtactgc
     4561 ggcaatcgtc aacttcactg gtttcaacgc ttcctagaag accaacagac taaacgcaaa
     4621 agcaaaaatc agcattctag cggcttgttc acactcagaa atggtcatct agtttggctt
     4681 gaaggtgagg gtaaagggga accttggaat cttcaccact tgacccttta ctgctgtgtt
     4741 gacaatcgct tgtggacaga ggagggaaca gaaatcgttc gccaagagaa agcagatgaa
     4801 attactaaat tcatcacaaa catgaagaag aaaagcgatc taagcgatac acagcaagct
     4861 ttgattcaac gtaaacaatc aacacttact cgaataaaca attcctttga gcgtcctagc
     4921 caaccccttt atcaaggtca atcacacatt ttggttggag taagcctggg actagaaaaa
     4981 cctgccacag tagcagtagt agatgcgatc gccaacaaag tcttggctta ccggagtatt
     5041 aaacaattac ttggcgacaa ttacgaactg ctaaatcgcc agagacgaca acagcagtac
     5101 ctatctcacg aacgccacaa agcacaaaaa aacttctctc ccaatcaatt tggagcatct
     5161 gagttagggc aacatataga cagattatta gctaaagcaa ttgtagcgtt agcgagaacc
     5221 tacaaagctg gcagtattgt cttgcccaag ttaggggata tgcgggaggt tgtccaaagt
     5281 gaaattcaag ctatagcaga acaaaaattt cccggttata ttgaaggtca gcaaaaatat
     5341 gccaaacagt accgggttaa tgttcatcgg tggagctacg gcagattaat tcaaagcatt
     5401 caaagtaaag cagctcaaac aggaattgtg attgaggagg gaaaacaacc tattcgaggt
     5461 agtccccacg acaaagcaaa ggaattagca ctttctgctt acaatctccg cctaactagg
     5521 cgaagttaac aaatatctga accttgataa tagaatatta atagcgccgc aattcatgct
     5581 gcttgcagcc tctgaatttt gttaaatgag ggttagtttg actgtataaa tacagtcttg
     5641 ctttctgacc ctggtagctg ctcaccctga tgctgctgtc aatagacagg ataggtgcgc
     5701 tcccagcaat aagggcgcgg atgtactgct gtagtggcta ctgaatcacc cccgatcaag
     5761 ggggaaccct ccccaattct tcatttgaag gactaaaatc aaggcaaaat ttctaagaga
     5821 tccgcgcaag ttccaaatac cttatctcgt cttgatttca tcttttttta accaaggcat
     5881 gattcttgag actgaggctc aaatgagaaa ttgggaaaca tccgcgctga agatatctgg
     5941 aaaggttggc tcacaacatt ttgtaactgg gtagttgaca gctagctcag tcctaggtat
     6001 aatgctagcg tggcaacaac cttccaggta ctaggtgggt tgaaaggaga agtcatttaa
     6061 taaggccact gttaaaagtg gcaacaacct tccaggtact aggtgggttg aaagcggccg
     6121 acgcgctggg ctactggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6181 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     6241 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     6301 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     6361 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     6421 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     6481 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     6541 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     6601 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     6661 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     6721 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     6781 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     6841 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     6901 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     6961 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     7021 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     7081 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     7141 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     7201 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     7261 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     7321 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     7381 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     7441 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     7501 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     7561 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     7621 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     7681 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     7741 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     7801 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     7861 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     7921 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     7981 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     8041 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     8101 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcc
//
