LOCUS       sequence-460482-        6969 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A single site gateway compatible destination vector with an upstream
            GLAST promotor to drive expression in glial cells. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6969)
  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 6969)
  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: pGLAST-DEST (JDW 1009)
FEATURES             Location/Qualifiers
     source          1..6969
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      129..584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(216..235)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     426..447
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    715..763
                     /note="synthetic polyadenylation signal"
     misc_feature    777..868
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     817..836
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    3021..3145
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3170..3175,3176..3193,3194..3200)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3254..3913
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3320..3339)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4255..4560
                     /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
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4439..4458
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4601..4725)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    4807..4928
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4818..4837)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4872..4891
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(5176..5193)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5347..5935)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5427..5446)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6106..6897,6898..6966))
                     /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     6729..6748
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(6967..6969,1..102))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
       61 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
      121 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      181 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      241 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      301 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      361 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      421 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      481 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      541 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt cgccattcag
      601 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa
      661 gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg ccgcaataaa
      721 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
      781 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
      841 gcaagtgcag gtgccagaac atttctctat cgataggtac catgtctaca caaactgaaa
      901 taatgaaatg gtttttaacc aagctttagg gattagagag gaaaaaaaaa gtgaaaaaga
      961 aagggcacca gggtcttccc atctctatgc taaccaaagt gtggttttcc agcccttttt
     1021 cttgcctatt aacaaaaata aaaacaaaaa atggaaatgc cactgggctt gaaaccccat
     1081 ttaaaatcag atttctcttt cgagttccct tgcaatacaa gtcaaagtca tttccaccaa
     1141 aaaaaaaaag cggtgggggg ggtgcgcaaa ttaatcctaa actcatggct cagataggac
     1201 ctggtatagt aaataaatgc ttttaaaagt agtcatggca ttgagaaatg gctccatgca
     1261 cctaaaccag aggcaccaat gtaaaaacta gtaattgtgt tttaacacaa aagattgaaa
     1321 agcgttttgt ttcacatcct aatattattc cacccactta aaaagagtta cttaatattg
     1381 actctgtatt aggtataaga caagttcttg ccctcaaaga acttataatt tagttgagaa
     1441 accaaaactt aacatgagct taaaaagaag gatgcagtac aaaacatggt ataattaagt
     1501 gctaaacaat gtagtataga tgctcagtga cctaggagat cagaggaagg gaagggtcac
     1561 tgagggctgg ggatgtcaaa agcatgtact ttcagtaagt tcttaaaatg atagcatgtt
     1621 agaaagctgt tataattgct gctgatttgt aattttaaag cgttgacatg attcaccatt
     1681 gctttccaac cagtagctgt ttaaaaggac atacagttga gggtgaaagg agatatttct
     1741 ttatgatctc cagagatttg tattgagggg aaaaaagaca tactttcttg ttgaataata
     1801 gaccaagaat tgagcatatt ttgcatgggg aatattaaat tcagtgaaca agaatccaaa
     1861 cctcaggctg tgattctggt ataaattttt gtttctaatg aaccattttc ttccttctaa
     1921 agaagattta agttttggga ataagtcacc tcaaaaaatt ttttttttat tttctaatat
     1981 ctattttcac aatttatttt tcttttaaga aaagtggact agaagctcaa gttaccttct
     2041 caaggcaatg agaatttttt agctctaaaa aagttgtgtg tggtagggag aacatttagc
     2101 atcctttaaa atggagacat ctgaacagca acctatcttt tgaatcgcca aatctcaagt
     2161 gtactttgca ctctcggtct taaaatgctc ttcattttct agtttggcaa aaattccagt
     2221 ctctttacat ttgctacctt ttccgtcttt gcaacacagg cagtgtgagc taaattaggt
     2281 ttgttaaaat gaagttttga agtttcaact gctgaaatag aggcatgtcc ctaactttag
     2341 acaaatttag agaatcgtag taagcattat tttttttcta catacttgtc tgcattcagt
     2401 gtcttcccaa gaaacgtcag ttaattcccc gattcgaaaa aggaactttc tcgtaacagt
     2461 tgtacaacca ggctaaagag cacaccctcg tcttccctga aagggaggaa acatgcaata
     2521 atgtgaggta attaaattta gccttagaaa agtctttgaa acgttctaat caagactttc
     2581 agctttccga ttgtgtgcgc tttatttttc aattcttcct agtcccggtt ttgcatttct
     2641 aatgacagtg aactgacaaa gcgtgaaagt ggtctaagga gcaaaacaaa gccagcaagc
     2701 ctgcttctgg tgtgccaaga ttaatcaata ttccagaaac ctcggggttt ccccctcctc
     2761 cctgtgcggt ttccattccc cactcctcct ttcccgggca cagactcctc cccttttctt
     2821 aagttgccct gatagtaact tgcagtttca gagcacatgc acactgtcag ggctagcctg
     2881 gctgcttacg cgcgctgcgg attgttgctc cgttgtacct gctggggaat tcacctcgtt
     2941 actgcttgat gggctcgaga tcctgttttg acctccatag aagacaccgg gaccgatcca
     3001 gcctccgcgg tggcggcatc acaagtttgt acaaaaaagc tgaacgagaa acgtaaaatg
     3061 atataaatat caatatatta aattagattt tgcataaaaa acagactaca taatactgta
     3121 aaacacaaca tatccagtca ctatggcggc cgcattaggc accccaggct ttacacttta
     3181 tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc
     3241 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     3301 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     3361 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     3421 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     3481 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     3541 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     3601 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     3661 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     3721 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     3781 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     3841 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     3901 gggcggggcg taaacgcgtg gatccggctt actaaaagcc agataacagt atgcgtattt
     3961 gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa
     4021 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg
     4081 ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag
     4141 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg
     4201 cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag
     4261 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     4321 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     4381 tcagataaag tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     4441 atgatgacca ccgatatggc cagtgtgccg gtctccgtta tcggggaaga agtggctgat
     4501 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     4561 atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg
     4621 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     4681 tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtgatgg ccgctctagc
     4741 gaagggatcc cccatcgaat gcatgcatct agagtcgggg cggccggccg cttcgagcag
     4801 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     4861 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     4921 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     4981 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga
     5041 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     5101 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     5161 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5221 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5281 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5341 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5401 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5461 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5521 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5581 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5641 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5701 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5761 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5821 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5881 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5941 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6001 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6061 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6121 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6181 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6241 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6301 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6361 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6421 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6481 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6541 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6601 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6661 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6721 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6781 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6841 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6901 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6961 actcatact
//
