LOCUS       sequence-458200-        7917 bp    DNA     circular SYN 15-APR-2025
DEFINITION  A CAGGS driven gateway compatible destination vector that contains 
            an IRES-myristoylated-TagBFP for labeling the cell membrane.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7917)
  AUTHORS   Gillespie W, Zhang Y, Ruiz OE, Cerda III J, Ortiz-Guzman J, Turner 
            WD, Largoza G, Sherman M, Mosser LE, Fujimoto E, Chien C-B, 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 2024.07.13.603267
REFERENCE   2  (bases 1 to 7917)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.07.13.603267"
COMMENT     Sequence Label: pCAGEN-DV-IRES-myr-BFP (JDW 491)
FEATURES             Location/Qualifiers
     source          1..7917
                     /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..3512)
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     misc_feature    join(3573..3581,3582..3584,3585..4159)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3746..3765)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     3975..3992
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4171..4188
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    4197..4321
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4362..4667)
                     /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(4464..4483)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(4987..5667)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     5582..5601
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(join(5721..5727,5728..5745,5746..5751))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5776..5900)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5941..5960)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5968..6984
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(6022..6044)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        7280..7659
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7690..7794
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7795..7863,7864..7917,1..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 ttaattaagc
     2821 ttgtgcccca gtttgctagg gaggtcgcag tatctggcca ctgccacctc gtgctgctcg
     2881 acgtaggtct cgttgttggc ctccttgatt ctttccagtc tgtagtccac atagtagacg
     2941 ccaggcatct tgaggttctt agcgggtttc ttggatctat atgtggtctt gatgtttgcg
     3001 atcagatggc tcccgcccac gagcttcagg gccatgtcgt ttctgccttc caggccgccg
     3061 tcagcggggt acagcgtctc ggtgaaggcc tcccagccga gtgttttctt ctgcatcaca
     3121 gggccgttgg atgtgaagtt cacccctctg atcttgacgt tgtagatgag gcagccgtcc
     3181 tggaggctgg tgtcctgggt agcggtcagc acgcccccgt cttcgtatgt ggtgactctc
     3241 tcccatgtga agccctcagg gaaggactgc ttgaagaagt cggggatgcc ctgggtgtgg
     3301 ttgatgaagg tcttgctgcc gtagaggaag ctagtagcca ggatgtcgaa ggcgaagggg
     3361 agagggccgc cctcgaccac cttgattctc atggtctggg tgccctcgta gggcttgcct
     3421 tcgccctcgg atgtgcactt gaagtgatgg ttgtccacgg tgccctccat gtacagcttc
     3481 atgtgcatgt tctccttaat cagctcgctc atgatctttt ggtcctcatc attcttttca
     3541 acctgtttgg ttcttctcat acagcacagc atggttgtgg ccatattatc atcgtgtttt
     3601 tcaaaggaaa accacgtccc cgtggttcgg ggggcctaga cgttttttta acctcgacta
     3661 aacacatgta aagcatgtgc accgaggccc cagatcagat cccatacaat ggggtacctt
     3721 ctgggcatcc ttcagcccct tgttgaatac gcttgaggag agccatttga ctctttccac
     3781 aactatccaa ctcacaacgt ggcactgggg ttgtgccgcc tttgcaggtg tatcttatac
     3841 acgtggcttt tggccgcaga ggcacctgtc gccaggtggg gggttccgct gcctgcaaag
     3901 ggtcgctaca gacgttgttt gtcttcaaga agcttccaga ggaactgctt ccttcacgac
     3961 attcaacaga ccttgcattc ctttggcgag aggggaaaga cccctaggaa tgctcgtcaa
     4021 gaagacaggg ccaggtttcc gggccctcac attgccaaaa gacggcaata tggtggaaaa
     4081 taacatatag acaaacgcac accggcctta ttccaagcgg cttcggccag taacgttagg
     4141 ggggggggag ggagaggggg gggcggaatt atttaggtga cactatagga tatcgaacca
     4201 ctttgtacaa gaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     4261 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat
     4321 ggtcgacctg cagactggct gtgtataagg gagcctgaca tttatattcc ccagaacatc
     4381 aggttaatgg cgtttttgat gtcattttcg cggtggctga gatcagccac ttcttccccg
     4441 ataacggaga ccggcacact ggccatatcg gtggtcatca tgcgccagct ttcatccccg
     4501 atatgcacca ccgggtaaag ttcacggggg actttatctg acagcagacg tgcactggcc
     4561 agggggatca ccatccgtcg cccgggcgtg tcaataatat cactctgtac atccacaaac
     4621 agacgataac ggctctctct tttataggtg taaaccttaa actgcatttc accagcccct
     4681 gttctcgtca gcaaaagagc cgttcatttc aataaaccgg gcgacctcag ccatcccttc
     4741 ctgattttcc gctttccagc gttcggcacg cagacgacgg gcttcattct gcatggttgt
     4801 gcttaccaga ccggagatat tgacatcata tatgccttga gcaactgata gctgtcgctg
     4861 tcaactgtca ctgtaatacg ctgcttcata gcatacctct ttttgacata cttcgggtat
     4921 acatatcagt atatattctt ataccgcaaa aatcagcgcg caaatacgca tactgttatc
     4981 tggcttttag taagccggat cctctagatt acgccccgcc tgccactcat cgcagtactg
     5041 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     5101 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     5161 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     5221 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     5281 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     5341 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     5401 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgga attccggatg
     5461 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     5521 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     5581 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     5641 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgacgg
     5701 atcctaactc aaaatccaca cattatacga gccggaagca taaagtgtaa agcctggggt
     5761 gcctaatgcg gccgccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt
     5821 tttttatgca aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca
     5881 gcttttttgt acaaacttgt ttgatatctc gaggaattct ttgccaaaat gatgagacag
     5941 cacaataacc agcacgttgc ccaggagctg taggaaaaag aagaaggcat gaacatggtt
     6001 agcagaggct ctagagccgc cggtcacacg ccagaagccg aaccccgccc tgccccgtcc
     6061 cccccgaagg cagccgtccc cccgcggaca gccccgaggc tggagaggga gaaggggacg
     6121 gcggcgcggc gacgcacgaa ggccctcccc gcccatttcc ttcctgccgg cgccgcaccg
     6181 cttcgccccg cgcccgctag agggggtgcg gcggcgcctc ccagatttcg gctccgcaca
     6241 gatttgggac aaaggaagtc cctgcgccct ctcgcacgat taccataaaa ggcaatggct
     6301 gcggctcgcc gcgcctcgac agccgccggc gctccggggg ccgccgcgcc cctcccccga
     6361 gccctccccg gcccgaggcg gccccgcccc gcccggcacc cccacctgcc gccacccccc
     6421 gcccggcacg gcgagccccg cgccacgccc cgtacggagc cccgcacccg aagccgggcc
     6481 gtgctcagca actcggggag gggggtgcag ggggggttgc agcccgaccg acgcgcccac
     6541 accccctgct caccccccca cgcacacacc ccgcacgcag cctttgttcc cctcgcagcc
     6601 ccccccgcac cgcggggcac cgcccccggc cgcgctcccc tcgcgcacac tgcggagcgc
     6661 acaaagcccc gcgccgcgcc cgcagcgctc acagccgccg ggcagcgcgg agccgcacgc
     6721 ggcgctcccc acgcacacac acacgcacgc accccccgag ccgctccccc cgcacaaagg
     6781 gccctcccgg agcccctcaa ggctttcacg cagccacaga aaagaaacaa gccgtcatta
     6841 aaccaagcgc taattacagc ccggaggaga agggccgtcc cgcccgctca cctgtgggag
     6901 taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg gcggcggagc ggggcacggg
     6961 gcgaaggcag cgcgcagcga ctcccgcccg ccgcgcgctt cgctttttat agggccgccg
     7021 ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg ccgctctgat tggctgccgc
     7081 cgcacctctc cgcctcgccc ccccccgccc ctcccccccc cccccccccc cccgcccccc
     7141 cccccccccc cccccccccc cccccccccc ccgcccccat cgctgcacaa aataattaaa
     7201 aaataaataa atacaaaatt gggggtgggg agggggggga gatggggaga gtgaagcaga
     7261 acgtggggct cacctcgacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     7321 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     7381 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     7441 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     7501 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     7561 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     7621 taattgatta ctattaataa ctagtcaata atcaatgtcg acccaggtgg cacttttcgg
     7681 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7741 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     7801 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     7861 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacga
//
