LOCUS       sequence-431487-        3322 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Fragmid fragment: (Selection) fluorescent reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3322)
  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 3322)
  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: AA279
FEATURES             Location/Qualifiers
     source          1..3322
                     /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"
     regulatory      444..453
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             450..1160
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     primer_bind     complement(1232..1248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1232..1248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1245..1267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1256..1272
                     /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(1280..1286,1287..1304,1305..1310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1325..1346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1463..1480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1634..2222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1714..1733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2400..3209)
                     /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     3123..3142
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3201..3220
                     /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 tgaagacgta ggagccacca tggtgtctaa gggcgaagag ctgattaagg
      481 agaacatgca catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca
      541 catccgaggg cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggtggtcg
      601 agggcggccc tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca
      661 gaaccttcat caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg
      721 gcttcacatg ggagagagtc accacatacg aggacggggg cgtgctgacc gctacccagg
      781 acaccagcct ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc
      841 catccaacgg ccctgtgatg cagaagaaaa cactcggctg ggaggccaac accgagatgc
      901 tgtaccccgc tgacggcggc ctggaaggca gaagcgacat ggccctgaag ctcgtgggcg
      961 ggggccacct gatctgcaac ttcaagacca catacagatc caagaaaccc gctaagaacc
     1021 tcaagatgcc cggcgtctac tatgtggacc acagactgga aagaatcaag gaggccgaca
     1081 aagagaccta cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca
     1141 aactggggca caaacttaat ggaagcacgt cttcatcgga tcccgggccc gtcgactgca
     1201 gaggcctgca tgcaagcttg gtgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1261 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1321 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1381 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1441 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1501 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1561 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1621 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1681 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1741 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1801 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1861 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1921 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1981 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2041 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2101 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2161 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2221 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2281 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2341 aatgaagttt taaatcaagc ccaatctgaa taatgttaca accaattaac caattctgat
     2401 tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata
     2461 ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat
     2521 aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct
     2581 attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact
     2641 gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc aacaggccag
     2701 ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc
     2761 gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa
     2821 tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat
     2881 tcttctaata cctggaatgc tgtttttccg gggatcgcag tggtgagtaa ccatgcatca
     2941 tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt
     3001 agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac
     3061 aactctggcg catcgggctt cccatacaag cgatagattg tcgcacctga ttgcccgaca
     3121 ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc
     3181 ctcgacgttt cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca
     3241 gacagtttta ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt
     3301 tgagacacgg gccagagctg ca
//
