LOCUS       sequence-431517-        3221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Cas protein) reverse TetR; binds to TRE in 
            presence of doxycycline.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3221)
  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 3221)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA391
FEATURES             Location/Qualifiers
     source          1..3221
                     /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"
     primer_bind     complement(1131..1147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1131..1147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1155..1171
                     /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(1179..1185,1186..1203,1204..1209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1224..1245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1362..1379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1533..2121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1613..1632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2299..3108)
                     /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     3022..3041
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3100..3119
                     /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 cagctctagg ctggacaaat ccaagatcat caatggggct
      481 ctggagttgc tgaacggcgt tgggattgag ggccttacta ctcgcaaact cgcacagaaa
      541 ctcggcgttg aacagccgac gctctactgg catgtgaaga acaaaagagc tctgctggac
      601 gcgcttccaa ttgagatgct cgaccgccat cacactcact catgcccgtt ggaacccgag
      661 tcctggcaag atttcctgcg gaacaatgca aagagctata gatgtgcatt gctgtctcat
      721 cgggatgggg ctaaggtgca tttgggaact cggcctaccg aaaagcaata cgaaaccctt
      781 gaaaaccagc tggcatttct gtgccaacag ggattctccc tggaaaatgc gctctacgcc
      841 ttgagtgcgg tcggccattt tactcttgga tgcgtactgg aagagcagga gcaccaagtt
      901 gccaaggaag aaagggaaac ccccaccaca gatagtatgc cgcctttgct gaagcaagcc
      961 atcgagctct tcgacaggca gggggcggaa ccagcttttc ttttcggcct ggagcttatc
     1021 atttgtgggt tggagaagca actcaagtgt gagagcgggg ggtccaagtc ttcatcggat
     1081 cccgggcccg tcgactgcag aggcctgcat gcaagcttgg tgtaatcatg gtcatagctg
     1141 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2221 tagatccttt taaattaaaa atgaagtttt aaatcaagcc caatctgaat aatgttacaa
     2281 ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact gcaatttatt
     2341 catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg aaggagaaaa
     2401 ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga ttccgactcg
     2461 tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat caagtgagaa
     2521 atcaccatga gtgacgactg aatccggtga gaatggcaaa agtttatgca tttctttcca
     2581 gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat caaccaaacc
     2641 gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt taaaaggaca
     2701 attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat caacaatatt
     2761 ttcacctgaa tcaggatatt cttctaatac ctggaatgct gtttttccgg ggatcgcagt
     2821 ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg gaagaggcat
     2881 aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg caacgctacc
     2941 tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaagc gatagattgt
     3001 cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat cagcatccat
     3061 gttggaattt aatcgcggcc tcgacgtttc ccgttgaata tggctcataa caccccttgt
     3121 attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc
     3181 aatgtaacat cagagatttt gagacacggg ccagagctgc a
//
