LOCUS       sequence-346789-        5405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-protein coding locus components from KraIscB-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5405)
  AUTHORS   Altae-Tran H, Kannan S, Demircioglu FE, Oshiro R, Nety SP, McKay LJ,
            Dlakic M, Inskeep WP, Makarova KS, Macrae RK, Koonin EV, Zhang F
  TITLE     The widespread IS200/IS605 transposon family encodes diverse 
            programmable RNA-guided endonucleases.
  JOURNAL   Science. 2021 Oct;374(6563):57-65. doi: 10.1126/science.abj6856. 
            Epub 2021 Sep 9.
  PUBMED    34591643
REFERENCE   2  (bases 1 to 5405)
  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.abj6856"; journalName: "Science"; date: "2021-10"; 
            volume: "374"; issue: "6563"; pages: "57-65"
COMMENT     Sequence Label: pHS0667 KraIscB-1 non-coding locus in pCOLADuet-1
FEATURES             Location/Qualifiers
     source          1..5405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      280..915
                     /direction=RIGHT
                     /label=ColA ori
                     /note="Plasmids containing the ColA origin of replication 
                     can be propagated in E. coli cells that contain additional 
                     plasmids with compatible origins."
     primer_bind     751..770
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        933..1024
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1025..1840
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1025..1840
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1098..1117)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2070..2117)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2113..2131
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2169..2213)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    2322..2346
                     /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)."
     promoter        complement(2347..2365)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2479..2496)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2552..2576
                     /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)."
     promoter        complement(2577..2595)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2641..2718
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(2738..2757)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     primer_bind     3273..3290
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3407..3428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3443..3448,3449..3466,3467..3473)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3481..3497
                     /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)."
     primer_bind     3486..3508
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3505..3521
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3505..3521
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4654..4671)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4654..4670)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4663..4685)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4879..4898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg
      121 caccgggatc tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
      181 cgcggggcat gactaacatg agaattacaa cttatatcgt atggggctga cttcaggtgc
      241 tacatttgaa gagataaatt gcactgaaat ctagagtgat ggtgtcggga atccgtaaag
      301 gatcttcttg agatcctttt acgatcgtcg taatctcctg ctctgtaaac gaaaaaaccg
      361 cctggggagg cggtttgatc gaaggttaag tcagttgggg aactgcttaa cctggtaact
      421 ggctttagtg gagcgcagat accaaatact gtcctttcag tgtagcctct gttaggccac
      481 cacttcaaga ctctcgatat ctaaatccac taattctcag ttaccaatgg ctgctgccag
      541 tggcgttttg tcgtgtcttt ccgggttgga ctcaagatga tagttaccgg ataaggcgca
      601 gcagtcgggc tgaacggggg gttcttgcac acagcccagc ttggagcgaa ctgtctacac
      661 ggaacgggac gtggtgattt gggtaaagcc tccaccacaa cacggacgcc gcaggacggg
      721 aacaggagag cgcaagaggg agccatcagg gggaaacgcc tggtatcttt atagtcctgt
      781 cgggtttcgc caccactgat ttgagcgtca gatttcgtga tgttcgtcag gggggcggag
      841 cctatggaaa aacggcttcg ctccggcctt attgtctctc tgctaagtat cctcctggca
      901 tcttctagga cgtttctgcg ctagcatgcc tatttgttta tttttctaaa tacattcaaa
      961 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     1021 gagtatgagc catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat
     1081 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     1141 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     1201 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     1261 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     1321 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     1381 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     1441 tccttttaac agcgaccgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg
     1501 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     1561 gaaagaaatg cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt
     1621 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     1681 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     1741 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     1801 taaattgcag tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccact tgcggagacc cggtcgtcag cttgtcgtcg gttcagggca gggtcgttaa
     1981 atagccgctt atgtctattg ctggtttacc ggtttattga ctaccggaag cagtgtgacc
     2041 gtgtgcttct caaatgcctg aggtttcagc aaaaaacccc tcaagacccg tttagaggcc
     2101 ccaaggggtt atgctagtta ttgctcagcg gtggcagcag cctaggttaa ttaagctgcg
     2161 ctagtagacg agtccatgtg ctggcgttca aatttcgcag cagcggtttc tttaccagac
     2221 tcgagggtac cgacgtcagc gatcgcgtgg ccggccgata tccaattgag atctgccata
     2281 tgtatatctc cttcttatac ttaactaata tactaagatg gggaattgtt atccgctcac
     2341 aattccccta tagtgagtcg tattaatttc gattatgcgg ccgtgtacaa tacgattact
     2401 ttctgttcga cttaagcatt atgcggccgc aagcttgtcg acctgcaggc gcgccgagct
     2461 cgaattcgga tcctggctgt ggtgatgatg gtgatggctg ctgcccatgg tatatctcct
     2521 tattaaagtt aaacaaaatt atttctacag gggaattgtt atccgctcac aattccccta
     2581 tagtgagtcg tattaatttc ctaatgcagg agtcgcataa gggagagcgt cgagatcccg
     2641 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     2701 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     2761 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     2821 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
     2881 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     2941 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     3001 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     3061 ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     3121 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     3181 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
     3241 gtcgcattgg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     3301 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     3361 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     3421 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     3481 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
     3541 catgcctgca gacgagtccc tggaggatcc tgcagacgag tccctggagg atcgatcctg
     3601 cagacgagtc cctggaggat cagcccacag gcacactgta tactccagaa gaagaggccg
     3661 cacccgtttg aggccgcacc aaatgtgaac taccactgag ctgaagacgc agtggcttct
     3721 tcggaagtca ctgaagacgc agaccaggag ctccttcgga agcttgagtt caccagactc
     3781 gtttccagaa atgggaacag cgttcgattg gtcatgacac ctgcggttga cgcatcagac
     3841 cgctgctctg tcgctgaggg ttaagtaggc ttgaggaaag ggccggtgct ctcagcgcaa
     3901 aaagcctttt gaacactgtc gagatgaagc cggattccct tcgtggtcac agcgaaggga
     3961 tacgcaccac ccggcgcttg ccggagcatt ttccgaaagg agttttctat gaacgtcgtc
     4021 tacgtcctct cgccggagag aacaccatta atgccctgtc aacctgccat tgcgaggttg
     4081 ttgctgaaac aaggaaaatc atcatggatg atgaaacaaa gaaccacgcc aaatccctca
     4141 ttctctatca cctcatcttt gtctgcaagt gcgggaaaaa acgtttgatg cgctctggag
     4201 atgagacaaa aaggatattc gaggtcattg caacacaatc agacgtttcc tttgagacga
     4261 tggaggtgga caaggatcat attcactgcc tagtaaaaag cgaaccgagg atagcaccac
     4321 tcgcgatagt acgaaggtta aagcaagaat cgactcttca gctttggcag atgcacgaag
     4381 atgaattgag aagacacttt tggaaggaaa ggacattctg gagcgatgga tacttctgcc
     4441 gtaccgttgg cgatgcaagt cagggaacaa gacgccagta cattgaggat caagggtgag
     4501 gtgcggcaat tcatccacga agctgaagac ttcgcggttt tcttgccgcc tttcctataa
     4561 aaaatggata caagcagggt gccttccctt gtaggaaggc accctgcttg tatccatcat
     4621 cttaccagcc agccgggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga
     4681 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     4741 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4801 tggcgaatgg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4861 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     4921 cccgccaaca cccgctgtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     4981 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     5041 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     5101 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     5161 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggacatctc ggtagtggga
     5221 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     5281 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     5341 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     5401 aatac
//
