LOCUS       sequence-255046-        8233 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral shuttle vector for shRNA expression from U6 promoter with
            GFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8233)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 8233)
  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"
COMMENT     Sequence Label: pShuttle-CMV-GFP U6promoter
FEATURES             Location/Qualifiers
     source          1..8233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        363..666
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        667..870
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     821..841
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      909..918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(915..917,918..920,921..1631)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(915..917,918..920,921..1631)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(960..981)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1221..1240)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1568..1589
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   4827..4929
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(5018..5035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5189..5777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5269..5288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6609..7403
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6609..7403
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6663..6682)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7273..7292
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7451..7831)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7513..7534)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7725..7744
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7869..7971
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     8059..8209
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg gaagataacc tgcttcaaca cgaatgttga agcaggttat cttccttttt
      301 tctagagatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      361 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      421 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      481 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      541 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      601 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      661 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      721 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      781 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      841 gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat ccgctagcgc
      901 taccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      961 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1021 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1081 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1141 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1201 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1261 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1321 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1381 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1441 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1501 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1561 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1621 agctgtacaa gtccggactc agatctgggc gtggttaagg gtgggaaaga atatataagg
     1681 tgggggtctt atgtagtttt gtatctgttt tgcagcagcc gccgccgcca tgagcaccaa
     1741 ctcgtttgat ggaagcattg tgagctcata tttgacaacg cgcatgcccc catgggccgg
     1801 ggtgcgtcag aatgtgatgg gctccagcat tgatggtcgc cccgtcctgc ccgcaaactc
     1861 tactaccttg acctacgaga ccgtgtctgg aacgccgttg gagactgcag cctccgccgc
     1921 cgcttcagcc gctgcagcca ccgcccgcgg gattgtgact gactttgctt tcctgagccc
     1981 gcttgcaagc agtgcagctt cccgttcatc cgcccgcgat gacaagttga cggctctttt
     2041 ggcacaattg gattctttga cccgggaact taatgtcgtt tctcagcagc tgttggatct
     2101 gcgccagcag gtttctgccc tgaaggcttc ctcccctccc aatgcggttt aaaacataaa
     2161 taaaaaacca gactctgttt ggatttggat caagcaagtg tcttgctgtc tttatttagg
     2221 ggttttgcgc gcgcggtagg cccgggacca gcggtctcgg tcgttgaggg tcctgtgtat
     2281 tttttccagg acgtggtaaa ggtgactctg gatgttcaga tacatgggca taagcccgtc
     2341 tctggggtgg aggtagcacc actgcagagc ttcatgctgc ggggtggtgt tgtagatgat
     2401 ccagtcgtag caggagcgct gggcgtggtg cctaaaaatg tctttcagta gcaagctgat
     2461 tgccaggggc aggcccttgg tgtaagtgtt tacaaagcgg ttaagctggg atgggtgcat
     2521 acgtggggat atgagatgca tcttggactg tatttttagg ttggctatgt tcccagccat
     2581 atccctccgg ggattcatgt tgtgcagaac caccagcaca gtgtatccgg tgcacttggg
     2641 aaatttgtca tgtagcttag aaggaaatgc gtggaagaac ttggagacgc ccttgtgacc
     2701 tccaagattt tccatgcatt cgtccataat gatggcaatg ggcccacggg cggcggcctg
     2761 ggcgaagata tttctgggat cactaacgtc atagttgtgt tccaggatga gatcgtcata
     2821 ggccattttt acaaagcgcg ggcggagggt gccagactgc ggtataatgg ttccatccgg
     2881 cccaggggcg tagttaccct cacagatttg catttcccac gctttgagtt cagatggggg
     2941 gatcatgtct acctgcgggg cgatgaagaa aacggtttcc ggggtagggg agatcagctg
     3001 ggaagaaagc aggttcctga gcagctgcga cttaccgcag ccggtgggcc cgtaaatcac
     3061 acctattacc ggctgcaact ggtagttaag agagctgcag ctgccgtcat ccctgagcag
     3121 gggggccact tcgttaagca tgtccctgac tcgcatgttt tccctgacca aatccgccag
     3181 aaggcgctcg ccgcccagcg atagcagttc ttgcaaggaa gcaaagtttt tcaacggttt
     3241 gagaccgtcc gccgtaggca tgcttttgag cgtttgacca agcagttcca ggcggtccca
     3301 cagctcggtc acctgctcta cggcatctcg atccagcata tctcctcgtt tcgcgggttg
     3361 gggcggcttt cgctgtacgg cagtagtcgg tgctcgtcca gacgggccag ggtcatgtct
     3421 ttccacgggc gcagggtcct cgtcagcgta gtctgggtca cggtgaaggg gtgcgctccg
     3481 ggctgcgcgc tggccagggt gcgcttgagg ctggtcctgc tggtgctgaa gcgctgccgg
     3541 tcttcgccct gcgcgtcggc caggtagcat ttgaccatgg tgtcatagtc cagcccctcc
     3601 gcggcgtggc ccttggcgcg cagcttgccc ttggaggagg cgccgcacga ggggcagtgc
     3661 agacttttga gggcgtagag cttgggcgcg agaaataccg attccgggga gtaggcatcc
     3721 gcgccgcagg ccccgcagac ggtctcgcat tccacgagcc aggtgagctc tggccgttcg
     3781 gggtcaaaaa ccaggtttcc cccatgcttt ttgatgcgtt tcttacctct ggtttccatg
     3841 agccggtgtc cacgctcggt gacgaaaagg ctgtccgtgt ccccgtatac agacttgaga
     3901 gggagtttaa acgaattcaa tagcttgttg catgggcggc gatataaaat gcaaggtgct
     3961 gctcaaaaaa tcaggcaaag cctcgcgcaa aaaagaaagc acatcgtagt catgctcatg
     4021 cagataaagg caggtaagct ccggaaccac cacagaaaaa gacaccattt ttctctcaaa
     4081 catgtctgcg ggtttctgca taaacacaaa ataaaataac aaaaaaacat ttaaacatta
     4141 gaagcctgtc ttacaacagg aaaaacaacc cttataagca taagacggac tacggccatg
     4201 ccggcgtgac cgtaaaaaaa ctggtcaccg tgattaaaaa gcaccaccga cagctcctcg
     4261 gtcatgtccg gagtcataat gtaagactcg gtaaacacat caggttgatt cacatcggtc
     4321 agtgctaaaa agcgaccgaa atagcccggg ggaatacata cccgcaggcg tagagacaac
     4381 attacagccc ccataggagg tataacaaaa ttaataggag agaaaaacac ataaacacct
     4441 gaaaaaccct cctgcctagg caaaatagca ccctcccgct ccagaacaac atacagcgct
     4501 tccacagcgg cagccataac agtcagcctt accagtaaaa aagaaaacct attaaaaaaa
     4561 caccactcga cacggcacca gctcaatcag tcacagtgta aaaaagggcc aagtgcagag
     4621 cgagtatata taggactaaa aaatgacgta acggttaaag tccacaaaaa acacccagaa
     4681 aaccgcacgc gaacctacgc ccagaaacga aagccaaaaa acccacaact tcctcaaatc
     4741 gtcacttccg ttttcccacg ttacgtcact tcccatttta agaaaactac aattcccaac
     4801 acatacaagt tactccgccc taaaacctac gtcacccgcc ccgttcccac gccccgcgcc
     4861 acgtcacaaa ctccaccccc tcattatcat attggcttca atccaaaata aggtatatta
     4921 ttgatgatgt taattaacat gcatggatcc atatgcggtg tgaaataccg cacagatgcg
     4981 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5041 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5101 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5161 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5221 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5281 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5341 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5401 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5461 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5521 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5581 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5641 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5701 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5761 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5821 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5881 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5941 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6001 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6061 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6121 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6181 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6241 tgcgcaacgt tgttgccatt gctgcagcca tgagattatc aaaaaggatc ttcacctaga
     6301 tccttttcac gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct
     6361 actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg
     6421 ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc
     6481 cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct
     6541 tgccgccaag gatctgatgg cgcaggggat caagctctga tcaagagaca ggatgaggat
     6601 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     6661 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     6721 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     6781 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     6841 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     6901 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     6961 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     7021 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     7081 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     7141 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     7201 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     7261 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     7321 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     7381 gccttcttga cgagttcttc tgaattttgt taaaattttt gttaaatcag ctcatttttt
     7441 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7501 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7561 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7621 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7681 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7741 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     7801 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggatc gaattaattc
     7861 ttaattaaca tcatcaataa tataccttat tttggattga agccaatatg ataatgaggg
     7921 ggtggagttt gtgacgtggc gcggggcgtg ggaacggggc gggtgacgta gtagtgtggc
     7981 ggaagtgtga tgttgcaagt gtggcggaac acatgtaagc gacggatgtg gcaaaagtga
     8041 cgtttttggt gtgcgccggt gtacacagga agtgacaatt ttcgcgcggt tttaggcgga
     8101 tgttgtagta aatttgggcg taaccgagta agatttggcc attttcgcgg gaaaactgaa
     8161 taagaggaag tgaaatctga ataattttgt gttactcata gcgcgtaata ctggtaccaa
     8221 ggtcgggcag gaa
//
