LOCUS       sequence-486871-        5693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway 3' entry clone containing an IRES Luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5693)
  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 5693)
  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: p3E-IRES-Luciferase-WPRE-bGH-pA (JDW 1509)
FEATURES             Location/Qualifiers
     source          1..5693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(624..747)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    join(749..1323,1324..1326,1327..1335)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(916..933)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1143..1162
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1336..2988
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1402..1420)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(3067..3087)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3485..3496)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3605..3621)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3606..3622)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(3623..3640)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3629..3740
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(3802..3897)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3934..3953)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3935..3953)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3958..3974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3958..3974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4076..4095)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4087..4896
                     /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(4154..4173)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      5043..5631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5532..5551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccctg cagctctaga gctcgaattc tacaggtcac
      601 taataccatc taagtagttg gttcatagtg actgcatatg ttgtgtttta cagtattatg
      661 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
      721 ctcgttcaac tttcttgtac aaagtggccc cctctccctc ccccccccct aacgttactg
      781 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
      841 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
      901 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
      961 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     1021 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     1081 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     1141 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     1201 gtatgggatc tgatctgggg cctcggtaca catgctttac atgtgtttag tcgaggttaa
     1261 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     1321 aatatggcca caaccatgga agacgccaaa aacataaaga aaggcccggc gccattctat
     1381 ccgctggaag atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg
     1441 gttcctggaa caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag
     1501 tacttcgaaa tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat
     1561 cacagaatcg tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg
     1621 ttatttatcg gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc
     1681 aacagtatgg gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa
     1741 attttgaacg tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa
     1801 acggattacc agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt
     1861 tttaatgaat acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc
     1921 atgaactcct ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc
     1981 tgcgtgagat tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact
     2041 gcgattttaa gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat
     2101 ttgatatgtg gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg
     2161 agccttcagg attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc
     2221 gccaaaagca ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt
     2281 ggcgctcccc tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt
     2341 atcaggcaag gatatgggct cactgagact acatcagcta ttctgattac acccgagggg
     2401 gatgataaac cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat
     2461 ctggataccg ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct
     2521 atgattatgt ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat
     2581 ggatggctac attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt
     2641 gaccgcctga agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa
     2701 tccatcttgc tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat
     2761 gacgccggtg aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa
     2821 aaagagatcg tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga
     2881 gttgtgtttg tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc
     2941 agagagatcc tcataaaggc caagaagggc ggaaagatcg ccgtgtaata atcgacacgc
     3001 gttaaggata cgtagtcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3061 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3121 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3181 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3241 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3301 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3361 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3421 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3481 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3541 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3601 gcatcgatac cgtcgacctc gacctcgact gtgccttcta gttgccagcc atctgttgtt
     3661 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3721 taaaatgagg aaattgcatc aacaacatct agacatggat cctatgaatt cggtctcgag
     3781 ggaggtgtgg gaggtttttt ccaactttat tatacaaagt tggcattata aaaaagcatt
     3841 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgga
     3901 gctccatggt agcgttaacg cggccgcgat atcccctata gtgagtcgta ttacatggtc
     3961 atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa
     4021 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     4081 ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat
     4141 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     4201 tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     4261 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     4321 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     4381 atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     4441 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     4501 tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg
     4561 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     4621 gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca
     4681 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     4741 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     4801 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     4861 ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa
     4921 cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc aaaatccctt
     4981 aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5041 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5101 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5161 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5221 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5281 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5341 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5401 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5461 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5521 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5581 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5641 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtt
//
