LOCUS       sequence-346790-        6223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Non-protein coding locus components from CRISPR-associated IscB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6223)
  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 6223)
  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: pHS0578 CRISPR associated IscB non-coding locu...
FEATURES             Location/Qualifiers
     source          1..6223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      680..1315
                     /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     1151..1170
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1333..1424
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1425..2240
                     /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             1425..2240
                     /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(1498..1517)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      complement(2470..2517)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2513..2531
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2569..2613)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     protein_bind    2722..2746
                     /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(2747..2765)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2879..2896)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2952..2976
                     /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(2977..2995)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        3041..3118
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(3138..3157)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
       61 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
      121 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
      181 ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa gacagctcat
      241 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
      301 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
      361 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
      421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      481 gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg
      541 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactaacatg
      601 agaattacaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
      661 gcactgaaat ctagagtgat ggtgtcggga atccgtaaag gatcttcttg agatcctttt
      721 acgatcgtcg taatctcctg ctctgtaaac gaaaaaaccg cctggggagg cggtttgatc
      781 gaaggttaag tcagttgggg aactgcttaa cctggtaact ggctttagtg gagcgcagat
      841 accaaatact gtcctttcag tgtagcctct gttaggccac cacttcaaga ctctcgatat
      901 ctaaatccac taattctcag ttaccaatgg ctgctgccag tggcgttttg tcgtgtcttt
      961 ccgggttgga ctcaagatga tagttaccgg ataaggcgca gcagtcgggc tgaacggggg
     1021 gttcttgcac acagcccagc ttggagcgaa ctgtctacac ggaacgggac gtggtgattt
     1081 gggtaaagcc tccaccacaa cacggacgcc gcaggacggg aacaggagag cgcaagaggg
     1141 agccatcagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc caccactgat
     1201 ttgagcgtca gatttcgtga tgttcgtcag gggggcggag cctatggaaa aacggcttcg
     1261 ctccggcctt attgtctctc tgctaagtat cctcctggca tcttctagga cgtttctgcg
     1321 ctagcatgcc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1381 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagc catattcaac
     1441 gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     1501 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
     1561 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     1621 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     1681 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag
     1741 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     1801 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgaccgcg
     1861 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     1921 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     1981 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2041 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2101 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2161 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2221 tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag
     2281 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccact tgcggagacc
     2341 cggtcgtcag cttgtcgtcg gttcagggca gggtcgttaa atagccgctt atgtctattg
     2401 ctggtttacc ggtttattga ctaccggaag cagtgtgacc gtgtgcttct caaatgcctg
     2461 aggtttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta
     2521 ttgctcagcg gtggcagcag cctaggttaa ttaagctgcg ctagtagacg agtccatgtg
     2581 ctggcgttca aatttcgcag cagcggtttc tttaccagac tcgagggtac cgacgtcagc
     2641 gatcgcgtgg ccggccgata tccaattgag atctgccata tgtatatctc cttcttatac
     2701 ttaactaata tactaagatg gggaattgtt atccgctcac aattccccta tagtgagtcg
     2761 tattaatttc gattatgcgg ccgtgtacaa tacgattact ttctgttcga cttaagcatt
     2821 atgcggccgc aagcttgtcg acctgcaggc gcgccgagct cgaattcgga tcctggctgt
     2881 ggtgatgatg gtgatggctg ctgcccatgg tatatctcct tattaaagtt aaacaaaatt
     2941 atttctacag gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc
     3001 ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa
     3061 aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt
     3121 gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc
     3181 ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc
     3241 gatggcggag ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc
     3301 gttgctgatt ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc
     3361 ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg
     3421 aagcggcgtc gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg
     3481 gctgatcatt aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac
     3541 taatgttccg gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt
     3601 ctcccatgaa gacggtacgc gactgggcgt ggagcatctg gtcgcattgg atgctgtcgg
     3661 aatggacgat gacagtagcc tgcaaatgat gacctggccg gaacgcgaac gtaatcgctg
     3721 gcggcgcttc ctgagtacca agaatcatcc ggtggacatt gtggcttttg cggagctccg
     3781 ctggctcttg gatgttcccg atccggtgat gccaacgaag gaagagatgg agcggaagta
     3841 taagcggcag gcacggtacg aacgggaaaa ggaaaggtgt tgcttgtgat gggctccaga
     3901 agccgattgt ttacatctgc acccactggc gttatattcg gcccgattga aggtggtggc
     3961 gctatactca ttttcggggg gttgtcagga aaaagttgga actcaaacca ctcctcccag
     4021 ttgtagcctc cactcatttt cggggtgctg tcaggtttaa cagtggcctt attaaatgac
     4081 ttctcgttgt agcctccact cattttcggg gtgctgtcag ggagtgcccg cgctaccctg
     4141 cttggatgct gggctggaag gcaaaaggcg ttcaaaaaaa tgggcggtgg ttgacagttt
     4201 tccggaaatg agtgcggatt ttttccgcag ctacaaccag actaagttct ttgggaacta
     4261 cgttattgtc gtcatcatac ccacgggtgc gtgccagcct gtggctctat ggtcgtgcat
     4321 taaacaggga ctttggaggg tgcccagtgt gtacggcgtc aaaagcggcg ataacattgt
     4381 cgaggcatac tttacgtgac ggcaacgctt cttcggaagt tgcgcccgat cgctagcttt
     4441 aaggctaatt caatgtccag agtcttggtt gttgacgctg atcgttgtcc gctggcgcct
     4501 tgtacgccaa gacgggctcg cctgctactg aacagtggca agtagacaaa gcatatgacc
     4561 aagctcttca attccgacgg ctatggctat ggctttctga aggctccaga gccacgccag
     4621 accgttatca gcgaatcctg aacgaaagcg ctacaaagag cccaatcgct cgtccaggcg
     4681 cttgattgct gcctgtgcct ctgccttgtg cagttgacgt tcagaacgtg cagcatcgag
     4741 ctgcgcttcc acacctgccg cctttgcaac cacttgcagg tctagacgta gttcgtcaat
     4801 ctcctggcgc aaccgctcta gctcttcgcc gacttccttg aatccgctga ttggcatttg
     4861 tttgaacccg ttattatgtt ttacaaaacc taatattggt tttgtgttgt cggtggtaag
     4921 tgatgagttc cactcgatct attagcgaac tcatagcgat gttgcccttg atggctacat
     4981 taggaatcta accaccagtc aattactggc ctgacaagtg ataacaaccc ataagggcaa
     5041 caaaaaaggc aacattcatt acgaaacaac tgttggctat attgttggct agtatgcctg
     5101 tttttgattt tatcccgctt gaatcctgac aattcgattc ctacggttag catcagacct
     5161 tcacgtgcgc ggcgttaaga agactgtttg aattcgcaac acccgccacc tgggccggta
     5221 gggcaaaaca agcgcaagcg atcaattagg ttcaggcgcg gttagccgcg ttctcagctg
     5281 cctcgatttc ttgccggtgc cttgccgccc aatctttcaa gcgactatcg gccttccttg
     5341 cttccagacg cccagcccgt acagcggcgt ccacatcgcc accacgctgg attcgcaaca
     5401 actgttcgcc tgcattgatc ggcagccgtc gcattgacaa tgcagctgct agtggcgctc
     5461 cacgtcgcaa ctcttccaat agcttggatt gctcaagcgg atctgcaaaa aacttcgcca
     5521 gtcttagctt ggattgctgt ttcataatcg accttacttt tgcctaacct tttaccttac
     5581 ttttgcctaa cgccagccta acgccagcct aaccgtgatg acccattgtg ccaacctaac
     5641 cttacctaac ctccccccgt agggggtaag gtaaggttgg gtaaggcatc aatccaggtt
     5701 aggtagcttc aaaactgatg tggtcagatc gcaaaaaata ttcatccctt gagccctcct
     5761 cacgaactag aaagacacct gcctccacca tcttgtcctt gagttgtttc ttggttttgt
     5821 cgtggggcgt gatacccgcc acctcaagcc ctttgtacca agcagctctg gtcacaatgc
     5881 cattttcgct ttgcgttctt agctcaaggt atagcgctct tgcttcctgc tgcttgcttg
     5941 ttagtgtctg agccttagtt ggttcagcga tttcgcattg ctcaagaatg caagactgga
     6001 cagggcttcc atcgtcttct gctagtggcc gccctagttc gtcgagaacg tcaaccggct
     6061 ccagtttgaa gcctatcggc gcaaacggct ctgcatcctt cgacttggtg caggttaggt
     6121 taatgaatcc tctaatcccg caagaggccc ggcagtcacc agcaaatcgc gctgttagcg
     6181 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gct
//
