LOCUS       sequence-447209-        4051 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible 5' entry clone containing the human EF1a 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4051)
  AUTHORS   Gillespie WB 3rd, Zhang Y, Ruiz OE, Cerda J 3rd, Ortiz-Guzman J, 
            Sherman M, Turner WD, Largoza G, Mosser LE, Fujimoto E, Chien CB, 
            Kwan KM, Arenkiel BR, Devine WP, Wythe JD
  TITLE     MultiSite Assembly of Gateway Induced Clones (MAGIC): a flexible 
            cloning toolbox for use in vertebrate model systems.
  JOURNAL   Development. 2025 Nov 15;152(22):dev204308. doi: 10.1242/dev.204308.
            Epub 2025 Nov 13.
  PUBMED    41230856
REFERENCE   2  (bases 1 to 4051)
  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: 
            "Development."; date: "2025-11-15"; volume: "152(22):dev204308. 
            doi"; pages: " 10.1242/dev.204308. Epub 2025 Nov 13"
COMMENT     Sequence Label: p5E-EF1a (JDW 1164)
FEATURES             Location/Qualifiers
     source          1..4051
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..86
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(245..272)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(364..450)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     499..521
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     513..530
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     514..530
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    570..665
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        667..685
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     710..726
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     711..727
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        816..1994
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1047..1985
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1942..1962
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(2002..2018)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2002..2018)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(2055..2073)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    2074..2197
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2270..2288)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2293..2309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2293..2309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2411..2430)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2422..3231
                     /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     complement(2489..2508)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3378..3966
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3867..3886
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
       61 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      121 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      181 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
      241 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
      301 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
      361 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
      421 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
      481 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
      541 ggcccgagtt aacgctacca tggagctcca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg ataagcaatg cttttttata atgccaactt tgtatagaaa
      661 agttggtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgaggtcgac
      721 ggtatcgata agctttgcaa agatggataa agttttaaac agagaggaat ctttgcagct
      781 aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt cagtgggcag
      841 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg
      901 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      961 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
     1021 gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc
     1081 tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg cagtacgtga
     1141 ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag
     1201 gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc
     1261 gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa
     1321 atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc
     1381 aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt
     1441 cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg
     1501 ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc
     1561 cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc
     1621 ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg
     1681 gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact
     1741 ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt
     1801 cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg
     1861 agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg
     1921 agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat
     1981 ttcaggtgtc gtgaggtacc ggatccacta gttctagagc ggccgccacc gcggtggagc
     2041 tccagctttt gttcccttta gtgagggtta attcaagttt gtacaaaaaa gttgaacgag
     2101 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
     2161 cataatactg taaaacacaa catatgcagt cactatgaac caactactta gatggtatta
     2221 gtgacctgta gaattcgagc tctagagctg cagggcggcc gcgatatccc ctatagtgag
     2281 tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca aaatctctga
     2341 tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata
     2401 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag gccgcgatta
     2461 aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa
     2521 tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt gtttctgaaa
     2581 catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg
     2641 acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg
     2701 ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga atatcctgat
     2761 tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct
     2821 gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga
     2881 atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt
     2941 gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc agtcgtcact
     3001 catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt
     3061 gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc
     3121 ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat
     3181 cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg
     3241 gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg cgcaagctca
     3301 tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga ccccgtagaa
     3361 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3421 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3481 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3541 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3601 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3661 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3721 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3781 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3841 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3901 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3961 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4021 cacatgttct ttcctgcgtt atcccctgat t
//
