LOCUS       sequence-432439-        2854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Pol 2 promoter) U1 small nuclear RNA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2854)
  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 2854)
  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: AA294
FEATURES             Location/Qualifiers
     source          1..2854
                     /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(764..780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(764..780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(777..799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    788..804
                     /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(812..818,819..836,837..842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    857..878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(995..1012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1166..1754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1246..1265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1932..2741)
                     /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     2655..2674
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2733..2752
                     /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 cacatggagg cggtactatg tagatgagaa ttcaggagca
      481 aactgggaaa agcaactgct tccaaatatt tgtgattttt acagtgtagt tttggaaaaa
      541 ctcttagcct accaattctt ctaagtgttt taaaatgtgg gagccagtac acatgaagtt
      601 atagagtgtt ttaatgaggc ttaaatattt accgtaacta tgaaatgcta cgcatatcat
      661 gctgttcagg ctccgtggcc acgcaactca tactaggagc gtcttcatcg gatcccgggc
      721 ccgtcgactg cagaggcctg catgcaagct tggtgtaatc atggtcatag ctgtttcctg
      781 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      841 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      901 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      961 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1021 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1081 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1141 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1201 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1261 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1321 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1381 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1441 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1501 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1561 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1621 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1681 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1741 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1801 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1861 ttttaaatta aaaatgaagt tttaaatcaa gcccaatctg aataatgtta caaccaatta
     1921 accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1981 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     2041 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     2101 tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     2161 tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt
     2221 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     2281 attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa
     2341 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct
     2401 gaatcaggat attcttctaa tacctggaat gctgtttttc cggggatcgc agtggtgagt
     2461 aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     2521 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     2581 tgtttcagaa acaactctgg cgcatcgggc ttcccataca agcgatagat tgtcgcacct
     2641 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2701 tttaatcgcg gcctcgacgt ttcccgttga atatggctca taacacccct tgtattactg
     2761 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
     2821 catcagagat tttgagacac gggccagagc tgca
//
