LOCUS       sequence-345505-        7178 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Endogenous locus of CRISPR-associated IscB (Chesapeake Bay) with Fn 
            spacer.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7178)
  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 7178)
  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: pHS0535 CRISPR-associated IscB (Chesapeake Bay...
FEATURES             Location/Qualifiers
     source          1..7178
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(1777..1782,1783..1799,1800..1805))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1832..1850)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1918..2022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2023..2091,2092..2883)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2241..2260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3054..3642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3543..3562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3796..3813
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3828..3968
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3894..3913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4054..4076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4070..4261)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5719..5738
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gtggaagacg caaccgcaag acccgctacc gtcctgcccg ttggctgaat cgcaagcgta
       61 ccggcccacc tgtgctatcc agtgctgaca cggtttctac gttggggaag tggctcgctc
      121 catcgctcca acatcgcatt gaagtcatca tgacctgggt gcatcgcttg cggcgctacc
      181 tgcccatcac ggcgatttca caagagattg tgcgctttga tatgcagaag atgcagaacc
      241 cggaaatcag tggcgtggag taccagcagg gcactctgtt cggttacgag gtgcgggaat
      301 acctgttgga caagtggcag cgacagtgcg gctattgcgg tgccaaggac aagcgtttgg
      361 aagtggatca cattgttcca cgtagccacg gcggcagtga tcgggtcagc aacctgacgc
      421 tatcttgtga accctgcaac aaaagaaaga accaacgtcc tgcggcagta ttcctggcaa
      481 aaaaaccgga agtgttgcag aaactccaac gtcaggccaa ggctccgctc aaggatgctg
      541 cagccgtcaa ctcaacccga tacgctctac tggaacgtct gaaggccact ggattgcctg
      601 tggaaattgc cagcggtggc cgcaccaagt tcaatcgttc tgagcgacag attcccaaga
      661 cacactggct ggatgccgct tgtgtcggag ccagtacacc ggaagtgctg cagtggaaag
      721 cagtcaagcc tttggcgatc aaggcgatgg gacacgggaa acggcaggtc gtcaacgtag
      781 atgcttacgg tttccctaga ggtaaggcca aaggaattcc ggttcatccg tttcgcaccg
      841 gcgacattgt tcgggctgaa attcccaagg gcaagtatgt cggcacctac gtcagtcgta
      901 ttgccgaaac tacaacatcg aagcctttag ctgggttcaa gtccaagact ggtaaacgca
      961 tacaatgcca cacaaagcat atgaccaagc tcttcaattc cgacggctat ggctatggct
     1021 ttctgaaggc tccagagcca cgccagaccg ttatcagcga atcctgaacg aaagcgctac
     1081 aaagagccca atcgctcatc cagacgcttg atcgctgttt gcgtctcagc cttatgcagt
     1141 tggcgttctg aacgtgcagc gtccagctgc gcttctacac ctgccgcctt tgcaactact
     1201 tgcaggtcta gacgtagttc gccaatctcc tggcgcaacc gctctagctc ttcgccgact
     1261 tccttgaatt cgctgattgg catttgtttg aacccgttat tatgttttgc aaaacctaat
     1321 attggttttg tgttgtcggt ggtaagtgat gagttccact cggaacgtta tcgaactcat
     1381 agcgatgttg cccttgatgg ctacgttagg aatctaatca ctagataatt actggcctgt
     1441 ccggtgataa cacttcttaa gggcaacaaa aagggcaaca ttcattagga acaattgact
     1501 ctggaaatca acgataacca acatcgcgct tgccttttaa ctacctccca gttgattctc
     1561 tttaggcttt tctggttgat cagcagccat tcaaaacaga agaaaatttc tctaatcaga
     1621 aataaaaatc ccgcaagagg cccggcagta ccggcataac caagcctatg cctacagcat
     1681 ccagggtgac ggtgccgagg atgacgatga gcgcattgtt agatttcata cacggtgcct
     1741 gactgcgtta gcaatttaac tgtgataaac taccgcatta aagcttatcg atgataagct
     1801 gtcaaacatg agaattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     1861 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     1921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2101 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2161 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2221 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     2281 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2341 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2401 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2461 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2521 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     2581 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2641 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2701 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2761 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2821 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2881 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2941 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3181 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3721 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     3781 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     3841 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     3901 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3961 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4021 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4081 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     4141 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     4201 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     4261 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     4321 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     4381 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     4441 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     4501 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     4561 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     4621 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     4681 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     4741 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     4801 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     4861 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     4921 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     4981 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     5041 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     5101 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     5161 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     5221 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     5281 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     5341 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     5401 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     5461 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     5521 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
     5581 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc
     5641 ggccacgggg cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg
     5701 agcccgatct tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc
     5761 gccggtgatg ccggccacga tgcgtccggc gtagaggatc cacaggacgg gtgtggtcgc
     5821 catgatcgcg tagtcgatag tggctccaag tagcgaagcg agcaggactg ggcggcggcc
     5881 aaagcggtcg gacagtgctc cgagaacggg tgcgcataga aattgcatca acgcatatag
     5941 cgctagcagc acgccatagt gactggcgat gctgtcggaa tggacgatgt ctatgcgatc
     6001 cattcactac cgccagatgc accagtgtcg atcattcagg agatgcttgc gcctccaagc
     6061 gctctacgcc gccgtcagcg ggctatgcag ctgctgaata ccagacgcca ccagaagctt
     6121 cagaaggccc gtcagtaggc gtataaaaag cactgaacgc aaccacatcc ctggccaaaa
     6181 cactgcacca caagccatct gcagcctacc tgtttacatc tgcacccact ggcgttatat
     6241 tcgtcggccc gatcgaagat gcggggtgct gtcagtttgg tcagtttggg ctgaacccaa
     6301 cggggtagtt gtagcctcca ctcattttcg gggtgctgtc aggctgatgc gtttagctca
     6361 ggcgaaagtc gatgttgtag cctccactca ttttcggggt gctgtcaggg tgggtggatt
     6421 catctggagt ctgggaatcg ttgtagcctc cactcatttt cggggtgctg tcaggtttaa
     6481 cagtggcctt attaaatgac ttctcgttgt agcctccact cattttcggg gtgctgtcag
     6541 gagtgcccgc gctaccctgc ttggatgctg ggctggaagg caaaaggcat tcaaaaaaat
     6601 gggcggtggt tgacagtttt ccggaaatga gtgcggattt tttccgcagc tacaaccaga
     6661 ctaagttctt tgggaactac gttattgtcg tcatcatacc cacgggtgcg tgccagcctg
     6721 tggctctatg gtcgtgcatt aaacagggac tttggagggt gcccagtgtg tacagcgtca
     6781 aaagcggcga taacattgtc gaggcatact ttacgtgacg gcaacgcttc ttcggaagtt
     6841 gcgcccgatc gctagcttta aggctaattc aatgtccaga gtcttggttg ttgatgcgaa
     6901 tcgttgtccg ctggcgcctt gtacgccaag acgggcccgc ttgctactga acagtggcaa
     6961 ggcggcggtg ttgcgtcgat acccattcac catcatcctc aaacaatcct accctactgc
     7021 aaacccacga cctgtgcggc tgaaactaga cccaggcagc aagactaccg gcattgccgt
     7081 agtcacagaa gccactggcg aagtggtctg ggcggcagaa ctccagcacc gtggtcagct
     7141 gatcaagaat gccctggaat cccgccgatc gctacgcc
//
