LOCUS       sequence-418046-        4908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express DpFnuc with pureexpress.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4908)
  AUTHORS   Jiang K, Lim J, Sgrizzi S, Trinh M, Kayabolen A, Yutin N, Bao W, 
            Kato K, Koonin EV, Gootenberg JS, Abudayyeh OO
  TITLE     Programmable RNA-guided DNA endonucleases are widespread in 
            eukaryotes and their viruses.
  JOURNAL   Sci Adv. 2023 Sep 29;9(39):eadk0171. doi: 10.1126/sciadv.adk0171. 
            Epub 2023 Sep 27.
  PUBMED    37756409
REFERENCE   2  (bases 1 to 4908)
  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: "Sci Adv."; 
            date: "2023-09-29"; volume: "9(39):eadk0171. doi"; pages: " 
            10.1126/sciadv.adk0171. Epub 2023 Sep 27"
COMMENT     Sequence Label: pKJ226
FEATURES             Location/Qualifiers
     source          1..4908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(143..162)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     262..284
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(322..340)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        408..512
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(513..581,582..1373)
                     /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(731..750)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1544..2132
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2033..2052
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2286..2303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2420..2441
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2456..2461,2462..2479,2480..2486)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2494..2510
                     /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     2499..2521
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2518..2534
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2518..2534
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(2555..2560,2561..2577,2578..2583)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(2949..2996)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2992..3010
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             4529..4551
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(4582..4601)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4583..4601)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4685..4704)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4824..4841)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4824..4840)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4833..4855)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
       61 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
      121 cgcatatatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
      181 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
      241 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
      301 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
      361 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
      421 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
      481 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
      541 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
      601 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
      661 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
      721 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
      781 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
      841 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
      901 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
      961 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1021 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1081 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1141 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1201 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1261 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1321 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1381 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1441 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     1501 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     1561 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1621 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1681 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1741 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     1801 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     1861 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     1921 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     1981 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2041 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2101 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2161 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2221 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2281 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     2341 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     2401 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     2461 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     2521 gaaacagcta tgacatgatt acgaattctc atgtttgaca gcttatcatc gataagcttt
     2581 aatgcggtag tttatcacag ttaaattgct aacgcagtca ggcaccgtgt atgaaatcta
     2641 acaatgcgct catcgtcatc ctcggcaccg tcaccctgga tgctgtaggc ataggcttgg
     2701 ttatgccggt actgccgggc ctcttgcggg atatcgtcca ttccgacagc atcgccagtc
     2761 actatggcgt gctgctagcg ctatatgcgt tgatgcaatt tctatgcgca cccgttctcg
     2821 gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct cgcttcgcta cttggagcca
     2881 ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg gatatccgga tatagttcct
     2941 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
     3001 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatct
     3061 cctaaggctc gagtcactgc tgttgtttct caaatgggaa catacgtaac gactttaaca
     3121 aaatgtttct agccccgttc acatcgcggt gggtgttata tttacactga tcgcagtaga
     3181 atgtctccga cttctccaag tttttgttaa tagtcccgca aactccgcaa gtcttgcttg
     3241 tataggcctc gttttgaacg tccaggatac actctgtatt tgtggttctg tgagttaaac
     3301 gctggcggaa acgatagtgt tgccagtata acaggtcacg tttagcgcgc ttacttaacg
     3361 gcttgggccg gtctgtcttg acaagttggg ccacgttaat tttgcccaaa gcgacatatt
     3421 gatattcctt aacaaggtgt gacgacaact tcaggtggat gtcatctaca aaagtcttca
     3481 acttcagatg agtcttcttt aatgatacta tgcgctcgcg cgcagcctgt tgttgttttt
     3541 tgttctgcgc tttcgtcaag tgcatgtggg cgtgatcgat cttgttatga agcttgctta
     3601 tcacatactg cttatcctcc tttatgccga cctggaatac gcagcaatca attggatcgt
     3661 agaccgtagc gaatgtgcgc ccgcctggat ctatcccgca aactctcttt tctatagtgt
     3721 cgttgctggc attgcgtcta gtatatttag ggtcacaagg gatgttcata ataaacattc
     3781 cgtcggctct cttaacgatc ttgacgtcat gatctatggg aggaatcttc gtaataggct
     3841 ttgcaagctt aagaaatctt tcttttgggt tgcttctctt ctcaaagttg tcaggcatca
     3901 tgcatatgta tctgttttgc atgttcgtgt tatctgtgct aacatctttg ctacttaagt
     3961 ggcgaatgaa caatttaggg acacaaaagc taccttggat gatgttactg acggacattg
     4021 gccgctttaa tttacttttc agagacttcg ccgacttata cgaggtacag aaattcttac
     4081 atgacgttaa ttttacactc gtcattttat tgttaaaaaa ccagtcatcg ttttccatac
     4141 gatattgcgg gtccacgtcg ttggcgttgg ttttacatac gattttctgc agttcaaatt
     4201 gatgaggttt caggcctttc tcgttaacca accaaaggca ccaattgtac gtgtaattcg
     4261 agacacgaag catttcattt aatacccgct tctgatgctt ggtgggtctg atcccaaccg
     4321 agaaagatgt taagaattta tctgactgcg tcttttgttt cttaacagag gccgattgta
     4381 ataactgaat gacatcacag tcggttttct cgtggttaac catgtccttg cgacggatat
     4441 attcccacca gtaccacatt gacttatgct tgtgaacgtt tgccgcatcc cacaaagtca
     4501 ggtcctcacg ctttcttttc atggtatatc tccttcttaa agttaaacaa aattatttct
     4561 agagggaaac cgttgtggtc tccctatagt gagtcgtatt aatttcgcgg gatcgagatc
     4621 tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     4681 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     4741 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     4801 atctccttgc atgcaagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4861 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgcc
//
