LOCUS       sequence-487087-        7896 bp    DNA     circular SYN 24-SEP-2025
DEFINITION  Gateway destination vector with a CAGGS promoter in the backbone as 
            well as an IRES-myr-mKate to label cell membranes. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7896)
  AUTHORS   Gillespie W, Zhang Y, Ruiz OE, Cerda J, Ortiz-Guzman J, Turner WD, 
            Largoza G, Sherman M, 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 with diverse applications in vertebrate model 
            systems.
  JOURNAL   bioRxiv [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267.
  PUBMED    39026881
REFERENCE   2  (bases 1 to 7896)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 24, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Jul 13:2024.07.13.603267. doi: 
            10.1101/2024.07.13.603267."
COMMENT     Sequence Label: pCAGEN-DV-IRES-myr-mKate (JDW 494)
FEATURES             Location/Qualifiers
     source          1..7896
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(96..115)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      909..1497
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1398..1417
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1651..1668
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    1736..1870
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1760..1779)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1814..1833
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1876..2071)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1890..2025
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2130..2151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2166..2171,2172..2189,2190..2196)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2204..2220
                     /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)."
     primer_bind     2209..2231
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2228..2244
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2228..2244
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2587..2606
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2605..2660
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2706..2725
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2811..3509)
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     misc_feature    join(3570..3578,3579..3581,3582..4155)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3742..3761)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     3971..3988
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4167..4184
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    4193..4317
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4358..4663)
                     /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     complement(4460..4479)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4983..5663)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5578..5597
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5717..5723,5724..5741,5742..5747))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5772..5896)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5937..5956)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5964..6980
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(6018..6040)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6981..7257)
                     /label=chicken beta-actin promoter
     enhancer        7259..7638
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7669..7773
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7774..7842,7843..7896,1..738)
                     /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"
ORIGIN
        1 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
       61 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      121 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      181 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      241 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
      301 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      361 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      421 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      481 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      541 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
      601 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      661 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      721 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      781 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
      841 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
      901 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
      961 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1021 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1081 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1141 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1201 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1261 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     1321 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     1381 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     1441 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     1501 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     1561 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1621 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1681 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagcggatcc
     1741 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     1801 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     1861 taaacaagtt agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     1921 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc
     1981 atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg
     2041 cgggactatg gttgctgact aattgagatg cggatccgct ggcacgacag gtttcccgac
     2101 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     2161 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     2221 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgggctgca ggtcgaggga
     2281 tctccataag agaagaggga cagctatgac tgggagtagt caggagagga ggaaaaatct
     2341 ggctagtaaa acatgtaagg aaaattttag ggatgttaaa gaaaaaaata acacaaaaca
     2401 aaatataaaa aaaatctaac ctcaagtcaa ggcttttcta tggaataagg aatggacagc
     2461 agggggctgt ttcatatact gatgacctct ttatagccaa cctttgttca tggcagccag
     2521 catatgggca tatgttgcca aactctaaac caaatactca ttctgatgtt ttaaatgatt
     2581 tgccctccca tatgtccttc cgagtgagag acacaaaaaa ttccaacaca ctattgcaat
     2641 gaaaataaat ttcctttatt agccagaagt cagatgctca aggggcttca tgatgtcccc
     2701 ataatttttg gcagagggaa aaagatctca gtggtatttg tgagccaggg cattggccac
     2761 accagccacc accttctgat aggcagcctg cacctgagga gtgcggccgc tcatctgtgc
     2821 cccagtttgc tagggaggtc gcagtatctg gccacagcca cctcgtgctg ctcgacgtag
     2881 gtctctttgt cggcctcctt gattctttcc agtcttctgt ccacatagta gacgccgggc
     2941 atcttgaggt tcttagcggg tttcttggat ctgtatgtgg tcttcaagtt gcagatcagg
     3001 tggcccccgc ccacgagctt cagggccatg tcggctctgc cttccaggcc gccgtcagcg
     3061 gggtacaggg tctcggtgga ggcctcccag ccgagtgttt tcttctgcat cacagggccg
     3121 ttggatggga agttcacccc tctgatcttg acgttgtaga tgaggcagcc gtcctggagg
     3181 ctggtgtcct gggtagcggt cagcacgccc ccgtcttcgt atgtggtgac tctctcccat
     3241 gtgaagccct cggggaagga ctgcttaaag aagtcgggga tgccctgggt gtggttgatg
     3301 aaggttttgc tgccgtacat gaagctggta gccaggatgt cgaaggcgaa ggggagaggg
     3361 ccgccctcga ccgccttgat tctcatggtc tgggtgccct cgtagggctt gccttcgccc
     3421 tcggatgtgc acttgaagtg gtggttgttc acggtgccct ccatgtacag cttcatgtgc
     3481 atgttctcct taatcagctc gctcaccatg atcttttggt cctcatcatt cttttcaacc
     3541 tgtttggttc ttctcataca gcacagcatg gttgtggcca tattatcatc gtgtttttca
     3601 aaggaaaacc acgtccccgt ggttcggggg gcctagacgt ttttttaacc tcgactaaac
     3661 acatgtaaag catgtgaccg aggccccaga tcagatccca tacaatgggg taccttctgg
     3721 gcatccttca gccccttgtt gaatacgctt gaggagagcc atttgactct ttccacaact
     3781 atccaactca caacgtggca ctggggttgt gccgcctttg caggtgtatc ttatacacgt
     3841 ggcttttggc cgcagaggca cctgtcgcca ggtggggggt tccgctgcct gcaaagggtc
     3901 gctacagacg ttgtttgtct tcaagaagct tccagaggaa ctgcttcctt cacgacattc
     3961 aacagacctt gcattccttt ggcgagaggg gaaagacccc taggaatgct cgtcaagaag
     4021 acagggccag gtttccgggc cctcacattg ccaaaagacg gcaatatggt ggaaaataac
     4081 atatagacaa acgcacaccg gccttattcc aagcggcttc ggccagtaac gttagggggg
     4141 ggggagggag aggggggggc ggaattattt aggtgacact ataggatatc gaaccacttt
     4201 gtacaagaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     4261 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggtc
     4321 gacctgcaga ctggctgtgt ataagggagc ctgacattta tattccccag aacatcaggt
     4381 taatggcgtt tttgatgtca ttttcgcggt ggctgagatc agccacttct tccccgataa
     4441 cggagaccgg cacactggcc atatcggtgg tcatcatgcg ccagctttca tccccgatat
     4501 gcaccaccgg gtaaagttca cgggggactt tatctgacag cagacgtgca ctggccaggg
     4561 ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact ctgtacatcc acaaacagac
     4621 gataacggct ctctctttta taggtgtaaa ccttaaactg catttcacca gcccctgttc
     4681 tcgtcagcaa aagagccgtt catttcaata aaccgggcga cctcagccat cccttcctga
     4741 ttttccgctt tccagcgttc ggcacgcaga cgacgggctt cattctgcat ggttgtgctt
     4801 accagaccgg agatattgac atcatatatg ccttgagcaa ctgatagctg tcgctgtcaa
     4861 ctgtcactgt aatacgctgc ttcatagcat acctcttttt gacatacttc gggtatacat
     4921 atcagtatat attcttatac cgcaaaaatc agcgcgcaaa tacgcatact gttatctggc
     4981 ttttagtaag ccggatcctc tagattacgc cccgcctgcc actcatcgca gtactgttgt
     5041 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     5101 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     5161 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     5221 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     5281 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     5341 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     5401 acactatccc atatcaccag ctcaccgtct ttcattgcca tacggaattc cggatgagca
     5461 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     5521 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     5581 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     5641 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgacggatcc
     5701 taactcaaaa tccacacatt atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5761 aatgcggccg ccatagtgac tggatatgtt gtgttttaca gtattatgta gtctgttttt
     5821 tatgcaaaat ctaatttaat atattgatat ttatatcatt ttacgtttct cgttcagctt
     5881 ttttgtacaa acttgtttga tatctcgagg aattctttgc caaaatgatg agacagcaca
     5941 ataaccagca cgttgcccag gagctgtagg aaaaagaaga aggcatgaac atggttagca
     6001 gaggctctag agccgccggt cacacgccag aagccgaacc ccgccctgcc ccgtcccccc
     6061 cgaaggcagc cgtccccccg cggacagccc cgaggctgga gagggagaag gggacggcgg
     6121 cgcggcgacg cacgaaggcc ctccccgccc atttccttcc tgccggcgcc gcaccgcttc
     6181 gccccgcgcc cgctagaggg ggtgcggcgg cgcctcccag atttcggctc cgcacagatt
     6241 tgggacaaag gaagtccctg cgccctctcg cacgattacc ataaaaggca atggctgcgg
     6301 ctcgccgcgc ctcgacagcc gccggcgctc cgggggccgc cgcgcccctc ccccgagccc
     6361 tccccggccc gaggcggccc cgccccgccc ggcaccccca cctgccgcca ccccccgccc
     6421 ggcacggcga gccccgcgcc acgccccgta cggagccccg cacccgaagc cgggccgtgc
     6481 tcagcaactc ggggaggggg gtgcaggggg ggttgcagcc cgaccgacgc gcccacaccc
     6541 cctgctcacc cccccacgca cacaccccgc acgcagcctt tgttcccctc gcagcccccc
     6601 ccgcaccgcg gggcaccgcc cccggccgcg ctcccctcgc gcacactgcg gagcgcacaa
     6661 agccccgcgc cgcgcccgca gcgctcacag ccgccgggca gcgcggagcc gcacgcggcg
     6721 ctccccacgc acacacacac gcacgcaccc cccgagccgc tccccccgca caaagggccc
     6781 tcccggagcc cctcaaggct ttcacgcagc cacagaaaag aaacaagccg tcattaaacc
     6841 aagcgctaat tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac
     6901 gcggtcagtc agagccgggg cgggcggcgc gaggcggcgg cggagcgggg cacggggcga
     6961 aggcagcgcg cagcgactcc cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc
     7021 cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc tctgattggc tgccgccgca
     7081 cctctccgcc tcgccccgcc ccgcccctcg ccccgccccg ccccgcctgg cgcgcgcccc
     7141 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
     7201 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
     7261 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     7321 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     7381 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     7441 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     7501 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     7561 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     7621 tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     7681 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     7741 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     7801 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     7861 aaagtaaaag atgctgaaga tcagttgggt gcacga
//
