LOCUS       sequence-495448-        7920 bp    DNA     circular SYN 17-DEC-2025
DEFINITION  Gateway destination vector with a CAGGS promoter in the backbone as 
            well as an IRES-myr-EGFP to label cell membranes. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7920)
  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 7920)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 17, 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: pCAGEN-DV-IRES-myr-EGFP (JDW 495)
FEATURES             Location/Qualifiers
     source          1..7920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             536..841
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     720..739
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(882..1006)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1015..1032)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    join(1044..1618,1619..1621,1622..1630)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1211..1228)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1438..1457
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1691..1693,1694..1696,1697..2410)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1736..1757)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1997..2016)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2344..2365
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2496..2515)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2561..2616
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2615..2634)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2977..2993)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2977..2993)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2990..3012)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3001..3017
                     /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(3025..3031,3032..3049,3050..3055))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3070..3091
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3150..3345
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3196..3331
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3351..3485
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3388..3407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3442..3461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3553..3570)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3724..4312)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3804..3823)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4483..5274,5275..5343))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5106..5125
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5344..5448)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5479..5858
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5860..6138
                     /label=chicken beta-actin promoter
     intron          6139..7155
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7079..7101
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7163..7182
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    7223..7347
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(7372..7377,7378..7395,7396..7402)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             join(7456..7920,1..216)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(7522..7541)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
       61 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
      121 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
      181 caggcggggc gtaatctaga ggatccggct tactaaaagc cagataacag tatgcgtatt
      241 tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa
      301 aaagaggtat gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt
      361 gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa
      421 gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc
      481 gcccggttta ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca
      541 gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag
      601 tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct
      661 gtcagataaa gtcccccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg
      721 catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag aagtggctga
      781 tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata
      841 aatgtcaggc tcccttatac acagccagtc tgcaggtcga ccatagtgac tggatatgtt
      901 gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat atattgatat
      961 ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggttcga tatcctatag
     1021 tgtcacctaa ataattccgc ccccccctct ccctcccccc cccctaacgt tactggccga
     1081 agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg
     1141 tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg
     1201 ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt
     1261 cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac
     1321 cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca
     1381 aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg
     1441 ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg
     1501 ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa
     1561 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     1621 ggccacaacc atgctgtgct gtatgagaag aaccaaacag gttgaaaaga atgatgagga
     1681 ccaaaagatc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1741 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1801 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1861 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1921 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1981 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2041 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2101 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2161 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt
     2221 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2281 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2341 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2401 gtacaagtaa gcggccgcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
     2461 gtggccaatg ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat
     2521 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     2581 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2641 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     2701 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     2761 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     2821 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     2881 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     2941 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3001 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3061 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3121 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     3181 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3241 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3301 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     3361 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3421 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3481 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3541 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3601 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3661 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3721 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3781 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3841 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3901 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3961 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4021 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4081 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4141 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4201 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4261 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4321 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4381 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4441 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4501 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4561 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4621 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4681 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4741 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4801 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4861 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4921 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4981 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5041 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5101 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5161 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5221 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5281 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5341 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5401 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5461 aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg
     5521 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     5581 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     5641 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     5701 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     5761 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     5821 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
     5881 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
     5941 aattattttg tgcagcgatg ggggcggggg gggggggggg ggcgcgcgcc aggcggggcg
     6001 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
     6061 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
     6121 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
     6181 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
     6241 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
     6301 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
     6361 ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg
     6421 cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc
     6481 gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa
     6541 ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg
     6601 ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg
     6661 gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg
     6721 ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga ggggcgcggc
     6781 ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg
     6841 taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct
     6901 gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg
     6961 aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc
     7021 cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     7081 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     7141 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     7201 aaagaattcc tcgagatatc aaacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
     7261 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     7321 taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt
     7381 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga
     7441 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     7501 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     7561 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     7621 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
     7681 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     7741 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     7801 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     7861 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
//
