LOCUS       sequence-255047-        8224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral shuttle vector for shRNA expression from H1 promoter with
            GFP marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8224)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 8224)
  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 H1promoter
FEATURES             Location/Qualifiers
     source          1..8224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     enhancer        364..667
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        668..871
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     822..842
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      910..919
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(916..918,919..921,922..1632)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(916..918,919..921,922..1632)
                     /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(961..982)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1222..1241)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1569..1590
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   4828..4930
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(5019..5036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5190..5778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5270..5289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6610..7404
                     /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             6610..7404
                     /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(6664..6683)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7274..7293
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7452..7832)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7514..7535)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7726..7745
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   7870..7972
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     8060..8210
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accacgcgtc ccccctaagg ttaagtcgcc
      241 ctcgttcaag agacgagggc gacttaacct taggtttttg gaaatcgata ccgtcgcatg
      301 gtctagagat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      361 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      421 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      481 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      541 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      601 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      661 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      721 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      781 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      841 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
      901 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      961 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1021 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1081 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1141 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1201 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1261 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1321 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1381 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1441 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1501 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1561 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1621 gagctgtaca agtccggact cagatctggg cgtggttaag ggtgggaaag aatatataag
     1681 gtgggggtct tatgtagttt tgtatctgtt ttgcagcagc cgccgccgcc atgagcacca
     1741 actcgtttga tggaagcatt gtgagctcat atttgacaac gcgcatgccc ccatgggccg
     1801 gggtgcgtca gaatgtgatg ggctccagca ttgatggtcg ccccgtcctg cccgcaaact
     1861 ctactacctt gacctacgag accgtgtctg gaacgccgtt ggagactgca gcctccgccg
     1921 ccgcttcagc cgctgcagcc accgcccgcg ggattgtgac tgactttgct ttcctgagcc
     1981 cgcttgcaag cagtgcagct tcccgttcat ccgcccgcga tgacaagttg acggctcttt
     2041 tggcacaatt ggattctttg acccgggaac ttaatgtcgt ttctcagcag ctgttggatc
     2101 tgcgccagca ggtttctgcc ctgaaggctt cctcccctcc caatgcggtt taaaacataa
     2161 ataaaaaacc agactctgtt tggatttgga tcaagcaagt gtcttgctgt ctttatttag
     2221 gggttttgcg cgcgcggtag gcccgggacc agcggtctcg gtcgttgagg gtcctgtgta
     2281 ttttttccag gacgtggtaa aggtgactct ggatgttcag atacatgggc ataagcccgt
     2341 ctctggggtg gaggtagcac cactgcagag cttcatgctg cggggtggtg ttgtagatga
     2401 tccagtcgta gcaggagcgc tgggcgtggt gcctaaaaat gtctttcagt agcaagctga
     2461 ttgccagggg caggcccttg gtgtaagtgt ttacaaagcg gttaagctgg gatgggtgca
     2521 tacgtgggga tatgagatgc atcttggact gtatttttag gttggctatg ttcccagcca
     2581 tatccctccg gggattcatg ttgtgcagaa ccaccagcac agtgtatccg gtgcacttgg
     2641 gaaatttgtc atgtagctta gaaggaaatg cgtggaagaa cttggagacg cccttgtgac
     2701 ctccaagatt ttccatgcat tcgtccataa tgatggcaat gggcccacgg gcggcggcct
     2761 gggcgaagat atttctggga tcactaacgt catagttgtg ttccaggatg agatcgtcat
     2821 aggccatttt tacaaagcgc gggcggaggg tgccagactg cggtataatg gttccatccg
     2881 gcccaggggc gtagttaccc tcacagattt gcatttccca cgctttgagt tcagatgggg
     2941 ggatcatgtc tacctgcggg gcgatgaaga aaacggtttc cggggtaggg gagatcagct
     3001 gggaagaaag caggttcctg agcagctgcg acttaccgca gccggtgggc ccgtaaatca
     3061 cacctattac cggctgcaac tggtagttaa gagagctgca gctgccgtca tccctgagca
     3121 ggggggccac ttcgttaagc atgtccctga ctcgcatgtt ttccctgacc aaatccgcca
     3181 gaaggcgctc gccgcccagc gatagcagtt cttgcaagga agcaaagttt ttcaacggtt
     3241 tgagaccgtc cgccgtaggc atgcttttga gcgtttgacc aagcagttcc aggcggtccc
     3301 acagctcggt cacctgctct acggcatctc gatccagcat atctcctcgt ttcgcgggtt
     3361 ggggcggctt tcgctgtacg gcagtagtcg gtgctcgtcc agacgggcca gggtcatgtc
     3421 tttccacggg cgcagggtcc tcgtcagcgt agtctgggtc acggtgaagg ggtgcgctcc
     3481 gggctgcgcg ctggccaggg tgcgcttgag gctggtcctg ctggtgctga agcgctgccg
     3541 gtcttcgccc tgcgcgtcgg ccaggtagca tttgaccatg gtgtcatagt ccagcccctc
     3601 cgcggcgtgg cccttggcgc gcagcttgcc cttggaggag gcgccgcacg aggggcagtg
     3661 cagacttttg agggcgtaga gcttgggcgc gagaaatacc gattccgggg agtaggcatc
     3721 cgcgccgcag gccccgcaga cggtctcgca ttccacgagc caggtgagct ctggccgttc
     3781 ggggtcaaaa accaggtttc ccccatgctt tttgatgcgt ttcttacctc tggtttccat
     3841 gagccggtgt ccacgctcgg tgacgaaaag gctgtccgtg tccccgtata cagacttgag
     3901 agggagttta aacgaattca atagcttgtt gcatgggcgg cgatataaaa tgcaaggtgc
     3961 tgctcaaaaa atcaggcaaa gcctcgcgca aaaaagaaag cacatcgtag tcatgctcat
     4021 gcagataaag gcaggtaagc tccggaacca ccacagaaaa agacaccatt tttctctcaa
     4081 acatgtctgc gggtttctgc ataaacacaa aataaaataa caaaaaaaca tttaaacatt
     4141 agaagcctgt cttacaacag gaaaaacaac ccttataagc ataagacgga ctacggccat
     4201 gccggcgtga ccgtaaaaaa actggtcacc gtgattaaaa agcaccaccg acagctcctc
     4261 ggtcatgtcc ggagtcataa tgtaagactc ggtaaacaca tcaggttgat tcacatcggt
     4321 cagtgctaaa aagcgaccga aatagcccgg gggaatacat acccgcaggc gtagagacaa
     4381 cattacagcc cccataggag gtataacaaa attaatagga gagaaaaaca cataaacacc
     4441 tgaaaaaccc tcctgcctag gcaaaatagc accctcccgc tccagaacaa catacagcgc
     4501 ttccacagcg gcagccataa cagtcagcct taccagtaaa aaagaaaacc tattaaaaaa
     4561 acaccactcg acacggcacc agctcaatca gtcacagtgt aaaaaagggc caagtgcaga
     4621 gcgagtatat ataggactaa aaaatgacgt aacggttaaa gtccacaaaa aacacccaga
     4681 aaaccgcacg cgaacctacg cccagaaacg aaagccaaaa aacccacaac ttcctcaaat
     4741 cgtcacttcc gttttcccac gttacgtcac ttcccatttt aagaaaacta caattcccaa
     4801 cacatacaag ttactccgcc ctaaaaccta cgtcacccgc cccgttccca cgccccgcgc
     4861 cacgtcacaa actccacccc ctcattatca tattggcttc aatccaaaat aaggtatatt
     4921 attgatgatg ttaattaaca tgcatggatc catatgcggt gtgaaatacc gcacagatgc
     4981 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5041 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5101 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5281 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5341 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5401 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5461 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5521 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5581 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5641 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5701 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5761 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5821 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5881 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5941 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6001 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6061 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6121 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6181 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6241 ttgcgcaacg ttgttgccat tgctgcagcc atgagattat caaaaaggat cttcacctag
     6301 atccttttca cgtagaaagc cagtccgcag aaacggtgct gaccccggat gaatgtcagc
     6361 tactgggcta tctggacaag ggaaaacgca agcgcaaaga gaaagcaggt agcttgcagt
     6421 gggcttacat ggcgatagct agactgggcg gttttatgga cagcaagcga accggaattg
     6481 ccagctgggg cgccctctgg taaggttggg aagccctgca aagtaaactg gatggctttc
     6541 ttgccgccaa ggatctgatg gcgcagggga tcaagctctg atcaagagac aggatgagga
     6601 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6661 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6721 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6781 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6841 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6901 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6961 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     7021 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     7081 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     7141 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     7201 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     7261 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     7321 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     7381 cgccttcttg acgagttctt ctgaattttg ttaaaatttt tgttaaatca gctcattttt
     7441 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7501 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     7561 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     7621 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     7681 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     7741 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     7801 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggat cgaattaatt
     7861 cttaattaac atcatcaata atatacctta ttttggattg aagccaatat gataatgagg
     7921 gggtggagtt tgtgacgtgg cgcggggcgt gggaacgggg cgggtgacgt agtagtgtgg
     7981 cggaagtgtg atgttgcaag tgtggcggaa cacatgtaag cgacggatgt ggcaaaagtg
     8041 acgtttttgg tgtgcgccgg tgtacacagg aagtgacaat tttcgcgcgg ttttaggcgg
     8101 atgttgtagt aaatttgggc gtaaccgagt aagatttggc cattttcgcg ggaaaactga
     8161 ataagaggaa gtgaaatctg aataattttg tgttactcat agcgcgtaat actggtacca
     8221 attc
//
