LOCUS       sequence-431511-        3536 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Fragmid fragment: (Cas protein) Renilla luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3536)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 3536)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA349
FEATURES             Location/Qualifiers
     source          1..3536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             445..1374
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="TSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     primer_bind     complement(1446..1462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1446..1462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1459..1481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1470..1486
                     /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(join(1494..1500,1501..1518,1519..1524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1539..1560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1677..1694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1848..2436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1928..1947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2614..3423)
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3337..3356
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3415..3434
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagactgg cagcacttcg aaagtttatg atccagaaca aaggaaacgg
      481 atgataactg gtccgcagtg gtgggccaga tgtaaacaaa tgaatgttct tgattcattt
      541 attaattatt atgattcaga aaaacatgca gaaaatgctg ttattttttt acatggtaac
      601 gcggcctctt cttatttatg gcgacatgtt gtgccacata ttgagccagt agcgcggtgt
      661 attataccag accttattgg tatgggcaaa tcaggcaaat ctggtaatgg ttcttatagg
      721 ttacttgatc attacaaata tcttactgca tggtttgaac ttcttaattt accaaagaag
      781 atcatttttg tcggccatga ttggggtgct tgtttggcat ttcattatag ctatgagcat
      841 caagataaga tcaaagcaat agttcacgct gaaagtgtag tagatgtgat tgaatcatgg
      901 gatgaatggc ctgatattga agaagatatt gcgttgatca aatctgaaga aggagaaaaa
      961 atggttttgg agaataactt cttcgtggaa accatgttgc catcaaaaat catgagaaag
     1021 ttagaaccag aagaatttgc agcatatctt gaaccattca aagagaaagg tgaagttcgt
     1081 cgtccaacat tatcatggcc tcgtgaaatc ccgttagtaa aaggtggtaa acctgacgtt
     1141 gtacaaattg ttaggaatta taatgcttat ctacgtgcaa gtgatgattt accaaaaatg
     1201 tttattgaat cggacccagg attcttttcc aatgctattg ttgaaggtgc caagaagttt
     1261 cctaatactg aatttgtcaa agtaaaaggt ctgcattttt cgcaagaaga tgcacctgat
     1321 gaaatgggaa aatatatcaa atcgttcgtt gagcgagttc tcaaaaatga acaagggtcc
     1381 aagtcttcat cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag cttggtgtaa
     1441 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1501 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1561 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1621 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1681 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1741 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1801 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1861 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1921 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1981 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2041 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2101 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2161 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2221 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2281 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2341 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2401 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aagcccaatc
     2581 tgaataatgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     2641 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     2701 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     2761 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     2821 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagttt
     2881 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     2941 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     3001 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     3061 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     3121 tccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     3181 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     3241 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     3301 caagcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     3361 taaatcagca tccatgttgg aatttaatcg cggcctcgac gtttcccgtt gaatatggct
     3421 cataacaccc cttgtattac tgtttatgta agcagacagt tttattgttc atgatgatat
     3481 atttttatct tgtgcaatgt aacatcagag attttgagac acgggccaga gctgca
//
