LOCUS       sequence-431452-        3782 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (Pol 2 promoter) human EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3782)
  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 3782)
  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: AA065
FEATURES             Location/Qualifiers
     source          1..3782
                     /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..1622
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          675..1613
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1570..1590
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(1692..1708)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1692..1708)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1705..1727)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1716..1732
                     /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(1740..1746,1747..1764,1765..1770))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1785..1806
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1923..1940)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2094..2682)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2174..2193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2860..3669)
                     /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     3583..3602
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3661..3680
                     /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 cacggctccg gtgcccgtca gtgggcagag cgcacatcgc
      481 ccacagtccc cgagaagttg gggggagggg tcggcaattg atccggtgcc tagagaaggt
      541 ggcgcggggt aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg
      601 ggggagaacc gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg
      661 ccgccagaac acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt
      721 atggcccttg cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg
      781 agcttcgggt tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc
      841 ctcgtgcttg agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc
      901 accttcgcgc ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac
      961 ctgctgcgac gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca
     1021 ctggtatttc ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat
     1081 gttcggcgag gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag
     1141 ctggccggcc tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg
     1201 caaggctggc ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg
     1261 ctgcagggag ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca
     1321 cacaaaggaa aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc
     1381 gggcgccgtc caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt
     1441 ggggggaggg gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta
     1501 ggccagcttg gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt
     1561 ggttcattct caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt
     1621 gaaggagcgt cttcatcgga tcccgggccc gtcgactgca gaggcctgca tgcaagcttg
     1681 gtgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     1741 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     1801 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1861 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     1921 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     1981 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2041 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2101 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2161 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2221 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2281 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2341 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     2401 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     2461 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     2521 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     2581 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     2641 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2701 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     2761 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaagc
     2821 ccaatctgaa taatgttaca accaattaac caattctgat tagaaaaact catcgagcat
     2881 caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt gaaaaagccg
     2941 tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa gatcctggta
     3001 tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc cctcgtcaaa
     3061 aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg agaatggcaa
     3121 aagtttatgc atttctttcc agacttgttc aacaggccag ccattacgct cgtcatcaaa
     3181 atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga gacgaaatac
     3241 gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc gcaggaacac
     3301 tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata cctggaatgc
     3361 tgtttttccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac ggataaaatg
     3421 cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca tctcatctgt
     3481 aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg catcgggctt
     3541 cccatacaag cgatagattg tcgcacctga ttgcccgaca ttatcgcgag cccatttata
     3601 cccatataaa tcagcatcca tgttggaatt taatcgcggc ctcgacgttt cccgttgaat
     3661 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     3721 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacgg gccagagctg
     3781 ca
//
