LOCUS       sequence-431497-        3037 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Pol 2 promoter) Copper-induced expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3037)
  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 3037)
  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: AA367
FEATURES             Location/Qualifiers
     source          1..3037
                     /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"
     promoter        444..870
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     primer_bind     complement(947..963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(947..963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(960..982)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    971..987
                     /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(995..1001,1002..1019,1020..1025))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1040..1061
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1178..1195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1349..1937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1429..1448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2115..2924)
                     /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     2838..2857
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2916..2935
                     /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 cacaattcgt tgcaggacag gatgtggtgc ccgatgtgac
      481 tagctctttg ctgcaggccg tcctatcctc tggttccgat aagagaccca gaactccggc
      541 cccccaccgc ccaccgccac ccccatacat atgtggtacg caagtaagag tgcctgcgca
      601 tgccccatgt gccccaccaa gagctttgca tcccatacaa gtccccaaag tggagaaccg
      661 aaccaattct tcgcgggcag aacaaaagct tctgcacacg tgtccactcg aatttggagc
      721 cggccggcgt gtgcaaaaga ggtgaatcga acgaaagacc cgtgtgtaaa gccgcgtttc
      781 caaaatgtat aaaaccgaga gcatctggcc aatgtgcatc agttgtggtc agcagcaaaa
      841 tcaagtgaat catctcagtg caactaaagg ggggatcagg agcgtcttca tcggatcccg
      901 ggcccgtcga ctgcagaggc ctgcatgcaa gcttggtgta atcatggtca tagctgtttc
      961 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1021 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1081 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1141 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1201 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1261 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1321 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1381 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1441 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1501 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1561 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1621 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1681 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1741 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1801 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1861 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1921 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1981 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2041 tccttttaaa ttaaaaatga agttttaaat caagcccaat ctgaataatg ttacaaccaa
     2101 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata
     2161 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     2221 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     2281 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     2341 ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact
     2401 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     2461 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     2521 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     2581 cctgaatcag gatattcttc taatacctgg aatgctgttt ttccggggat cgcagtggtg
     2641 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     2701 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     2761 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaagcgata gattgtcgca
     2821 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     2881 gaatttaatc gcggcctcga cgtttcccgt tgaatatggc tcataacacc ccttgtatta
     2941 ctgtttatgt aagcagacag ttttattgtt catgatgata tatttttatc ttgtgcaatg
     3001 taacatcaga gattttgaga cacgggccag agctgca
//
