LOCUS       sequence-255181-        7932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral shuttle vector with CMV promoter-EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7932)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 7932)
  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
FEATURES             Location/Qualifiers
     source          1..7932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   9..111
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     199..349
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        435..738
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        739..942
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     893..913
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      981..990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(987..989,990..992,993..1703)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(987..989,990..992,993..1703)
                     /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(1032..1053)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1293..1312)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1640..1661
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     repeat_region   4899..5001
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     primer_bind     complement(5090..5107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5261..5849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5341..5360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             6681..7475
                     /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             6681..7475
                     /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(6735..6754)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7345..7364
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      complement(7523..7903)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7585..7606)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7797..7816
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttaattaaca tcatcaataa tataccttat tttggattga agccaatatg ataatgaggg
       61 ggtggagttt gtgacgtggc gcggggcgtg ggaacggggc gggtgacgta gtagtgtggc
      121 ggaagtgtga tgttgcaagt gtggcggaac acatgtaagc gacggatgtg gcaaaagtga
      181 cgtttttggt gtgcgccggt gtacacagga agtgacaatt ttcgcgcggt tttaggcgga
      241 tgttgtagta aatttgggcg taaccgagta agatttggcc attttcgcgg gaaaactgaa
      301 taagaggaag tgaaatctga ataattttgt gttactcata gcgcgtaata ctggtaccgc
      361 ggccgcctcg agtctagaga ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      421 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      481 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      541 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      601 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      661 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      721 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      781 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      841 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      901 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      961 atccgctagc gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     1021 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1081 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     1141 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     1201 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1261 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1321 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1381 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1441 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     1501 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1561 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     1621 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1681 tcggcatgga cgagctgtac aagtccggac tcagatctgg gcgtggttaa gggtgggaaa
     1741 gaatatataa ggtgggggtc ttatgtagtt ttgtatctgt tttgcagcag ccgccgccgc
     1801 catgagcacc aactcgtttg atggaagcat tgtgagctca tatttgacaa cgcgcatgcc
     1861 cccatgggcc ggggtgcgtc agaatgtgat gggctccagc attgatggtc gccccgtcct
     1921 gcccgcaaac tctactacct tgacctacga gaccgtgtct ggaacgccgt tggagactgc
     1981 agcctccgcc gccgcttcag ccgctgcagc caccgcccgc gggattgtga ctgactttgc
     2041 tttcctgagc ccgcttgcaa gcagtgcagc ttcccgttca tccgcccgcg atgacaagtt
     2101 gacggctctt ttggcacaat tggattcttt gacccgggaa cttaatgtcg tttctcagca
     2161 gctgttggat ctgcgccagc aggtttctgc cctgaaggct tcctcccctc ccaatgcggt
     2221 ttaaaacata aataaaaaac cagactctgt ttggatttgg atcaagcaag tgtcttgctg
     2281 tctttattta ggggttttgc gcgcgcggta ggcccgggac cagcggtctc ggtcgttgag
     2341 ggtcctgtgt attttttcca ggacgtggta aaggtgactc tggatgttca gatacatggg
     2401 cataagcccg tctctggggt ggaggtagca ccactgcaga gcttcatgct gcggggtggt
     2461 gttgtagatg atccagtcgt agcaggagcg ctgggcgtgg tgcctaaaaa tgtctttcag
     2521 tagcaagctg attgccaggg gcaggccctt ggtgtaagtg tttacaaagc ggttaagctg
     2581 ggatgggtgc atacgtgggg atatgagatg catcttggac tgtattttta ggttggctat
     2641 gttcccagcc atatccctcc ggggattcat gttgtgcaga accaccagca cagtgtatcc
     2701 ggtgcacttg ggaaatttgt catgtagctt agaaggaaat gcgtggaaga acttggagac
     2761 gcccttgtga cctccaagat tttccatgca ttcgtccata atgatggcaa tgggcccacg
     2821 ggcggcggcc tgggcgaaga tatttctggg atcactaacg tcatagttgt gttccaggat
     2881 gagatcgtca taggccattt ttacaaagcg cgggcggagg gtgccagact gcggtataat
     2941 ggttccatcc ggcccagggg cgtagttacc ctcacagatt tgcatttccc acgctttgag
     3001 ttcagatggg gggatcatgt ctacctgcgg ggcgatgaag aaaacggttt ccggggtagg
     3061 ggagatcagc tgggaagaaa gcaggttcct gagcagctgc gacttaccgc agccggtggg
     3121 cccgtaaatc acacctatta ccggctgcaa ctggtagtta agagagctgc agctgccgtc
     3181 atccctgagc aggggggcca cttcgttaag catgtccctg actcgcatgt tttccctgac
     3241 caaatccgcc agaaggcgct cgccgcccag cgatagcagt tcttgcaagg aagcaaagtt
     3301 tttcaacggt ttgagaccgt ccgccgtagg catgcttttg agcgtttgac caagcagttc
     3361 caggcggtcc cacagctcgg tcacctgctc tacggcatct cgatccagca tatctcctcg
     3421 tttcgcgggt tggggcggct ttcgctgtac ggcagtagtc ggtgctcgtc cagacgggcc
     3481 agggtcatgt ctttccacgg gcgcagggtc ctcgtcagcg tagtctgggt cacggtgaag
     3541 gggtgcgctc cgggctgcgc gctggccagg gtgcgcttga ggctggtcct gctggtgctg
     3601 aagcgctgcc ggtcttcgcc ctgcgcgtcg gccaggtagc atttgaccat ggtgtcatag
     3661 tccagcccct ccgcggcgtg gcccttggcg cgcagcttgc ccttggagga ggcgccgcac
     3721 gaggggcagt gcagactttt gagggcgtag agcttgggcg cgagaaatac cgattccggg
     3781 gagtaggcat ccgcgccgca ggccccgcag acggtctcgc attccacgag ccaggtgagc
     3841 tctggccgtt cggggtcaaa aaccaggttt cccccatgct ttttgatgcg tttcttacct
     3901 ctggtttcca tgagccggtg tccacgctcg gtgacgaaaa ggctgtccgt gtccccgtat
     3961 acagacttga gagggagttt aaacgaattc aatagcttgt tgcatgggcg gcgatataaa
     4021 atgcaaggtg ctgctcaaaa aatcaggcaa agcctcgcgc aaaaaagaaa gcacatcgta
     4081 gtcatgctca tgcagataaa ggcaggtaag ctccggaacc accacagaaa aagacaccat
     4141 ttttctctca aacatgtctg cgggtttctg cataaacaca aaataaaata acaaaaaaac
     4201 atttaaacat tagaagcctg tcttacaaca ggaaaaacaa cccttataag cataagacgg
     4261 actacggcca tgccggcgtg accgtaaaaa aactggtcac cgtgattaaa aagcaccacc
     4321 gacagctcct cggtcatgtc cggagtcata atgtaagact cggtaaacac atcaggttga
     4381 ttcacatcgg tcagtgctaa aaagcgaccg aaatagcccg ggggaataca tacccgcagg
     4441 cgtagagaca acattacagc ccccatagga ggtataacaa aattaatagg agagaaaaac
     4501 acataaacac ctgaaaaacc ctcctgccta ggcaaaatag caccctcccg ctccagaaca
     4561 acatacagcg cttccacagc ggcagccata acagtcagcc ttaccagtaa aaaagaaaac
     4621 ctattaaaaa aacaccactc gacacggcac cagctcaatc agtcacagtg taaaaaaggg
     4681 ccaagtgcag agcgagtata tataggacta aaaaatgacg taacggttaa agtccacaaa
     4741 aaacacccag aaaaccgcac gcgaacctac gcccagaaac gaaagccaaa aaacccacaa
     4801 cttcctcaaa tcgtcacttc cgttttccca cgttacgtca cttcccattt taagaaaact
     4861 acaattccca acacatacaa gttactccgc cctaaaacct acgtcacccg ccccgttccc
     4921 acgccccgcg ccacgtcaca aactccaccc cctcattatc atattggctt caatccaaaa
     4981 taaggtatat tattgatgat gttaattaac atgcatggat ccatatgcgg tgtgaaatac
     5041 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     5101 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5161 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5221 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5281 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5341 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5401 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5461 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5521 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5581 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5641 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5701 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5761 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     5821 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5881 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5941 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6001 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     6061 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     6121 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     6181 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     6241 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     6301 cagttaatag tttgcgcaac gttgttgcca ttgctgcagc catgagatta tcaaaaagga
     6361 tcttcaccta gatccttttc acgtagaaag ccagtccgca gaaacggtgc tgaccccgga
     6421 tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag agaaagcagg
     6481 tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg acagcaagcg
     6541 aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc aaagtaaact
     6601 ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagctct gatcaagaga
     6661 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     6721 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     6781 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     6841 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     6901 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     6961 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     7021 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     7081 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     7141 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     7201 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     7261 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     7321 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     7381 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     7441 tcgccttcta tcgccttctt gacgagttct tctgaatttt gttaaaattt ttgttaaatc
     7501 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     7561 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     7621 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     7681 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     7741 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     7801 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     7861 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg ccattcagga
     7921 tcgaattaat tc
//
