LOCUS       sequence-254862-        8971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Adenoviral shuttle vector with CMV promoter, coexpresses EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8971)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 8971)
  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-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..115
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     rep_origin      153..533
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(240..259)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     450..471
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(581..1375)
                     /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             complement(581..1375)
                     /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(692..711)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1302..1321
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     rep_origin      2207..2795
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2696..2715
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2949..2966
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   3055..3157
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     enhancer        6416..6719
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6720..6923
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6874..6894
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    6951..7007
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    join(7009..7583,7584..7586,7587..7595)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7176..7193)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7403..7422
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7596..7598,7599..7601,7602..8315)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(7596..7598,7599..7601,7602..8312)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7641..7662)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7902..7921)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8249..8270
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    8482..8603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8519..8538)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8573..8592
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_signal     8746..8896
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
ORIGIN
        1 tccgccacac tactacgtca cccgccccgt tcccacgccc cgcgccacgt cacaaactcc
       61 accccctcat tatcatattg gcttcaatcc aaaataaggt atattattga tgatgttaat
      121 taagaattaa ttcgatcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
      181 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
      241 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
      301 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
      361 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
      421 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
      481 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
      541 gttaaaaaat gagctgattt aacaaaaatt ttaacaaaat tcagaagaac tcgtcaagaa
      601 ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc
      661 ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct
      721 gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt
      781 ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg
      841 gcatgctcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt
      901 ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat
      961 gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg
     1021 catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc
     1081 ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag
     1141 ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcttgcagtt
     1201 cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca
     1261 gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata
     1321 gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa
     1381 acgatcctca tcctgtctct tgatcagagc ttgatcccct gcgccatcag atccttggcg
     1441 gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag
     1501 ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
     1561 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
     1621 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtgaaaa
     1681 ggatctaggt gaagatcctt tttgataatc tcatggctgc agcaatggca acaacgttgc
     1741 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     1801 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     1861 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     1921 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     1981 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2041 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2101 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2161 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     2221 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     2281 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     2341 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     2401 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     2461 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     2521 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     2581 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     2641 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     2701 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     2761 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     2821 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     2881 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     2941 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3001 ttacgcatct gtgcggtatt tcacaccgca tatggatcca tgcatgttaa ttaacatcat
     3061 caataatata ccttattttg gattgaagcc aatatgataa tgagggggtg gagtttgtga
     3121 cgtggcgcgg ggcgtgggaa cggggcgggt gacgtaggtt ttagggcgga gtaacttgta
     3181 tgtgttggga attgtagttt tcttaaaatg ggaagtgacg taacgtggga aaacggaagt
     3241 gacgatttga ggaagttgtg ggttttttgg ctttcgtttc tgggcgtagg ttcgcgtgcg
     3301 gttttctggg tgttttttgt ggactttaac cgttacgtca ttttttagtc ctatatatac
     3361 tcgctctgca cttggccctt ttttacactg tgactgattg agctggtgcc gtgtcgagtg
     3421 gtgttttttt aataggtttt cttttttact ggtaaggctg actgttatgg ctgccgctgt
     3481 ggaagcgctg tatgttgttc tggagcggga gggtgctatt ttgcctaggc aggagggttt
     3541 ttcaggtgtt tatgtgtttt tctctcctat taattttgtt atacctccta tgggggctgt
     3601 aatgttgtct ctacgcctgc gggtatgtat tcccccgggc tatttcggtc gctttttagc
     3661 actgaccgat gtgaatcaac ctgatgtgtt taccgagtct tacattatga ctccggacat
     3721 gaccgaggag ctgtcggtgg tgctttttaa tcacggtgac cagttttttt acggtcacgc
     3781 cggcatggcc gtagtccgtc ttatgcttat aagggttgtt tttcctgttg taagacaggc
     3841 ttctaatgtt taaatgtttt tttgttattt tattttgtgt ttatgcagaa acccgcagac
     3901 atgtttgaga gaaaaatggt gtctttttct gtggtggttc cggagcttac ctgcctttat
     3961 ctgcatgagc atgactacga tgtgctttct tttttgcgcg aggctttgcc tgattttttg
     4021 agcagcacct tgcattttat atcgccgccc atgcaacaag ctattgaatt cgtttaaact
     4081 ccctctcaag tctgtatacg gggacacgga cagccttttc gtcaccgagc gtggacaccg
     4141 gctcatggaa accagaggta agaaacgcat caaaaagcat gggggaaacc tggtttttga
     4201 ccccgaacgg ccagagctca cctggctcgt ggaatgcgag accgtctgcg gggcctgcgg
     4261 cgcggatgcc tactccccgg aatcggtatt tctcgcgccc aagctctacg ccctcaaaag
     4321 tctgcactgc ccctcgtgcg gcgcctcctc caagggcaag ctgcgcgcca agggccacgc
     4381 cgcggagggg ctggactatg acaccatggt caaatgctac ctggccgacg cgcagggcga
     4441 agaccggcag cgcttcagca ccagcaggac cagcctcaag cgcaccctgg ccagcgcgca
     4501 gcccggagcg caccccttca ccgtgaccca gactacgctg acgaggaccc tgcgcccgtg
     4561 gaaagacatg accctggccc gtctggacga gcaccgacta ctgccgtaca gcgaaagccg
     4621 ccccaacccg cgaaacgagg agatatgctg gatcgagatg ccgtagagca ggtgaccgag
     4681 ctgtgggacc gcctggaact gcttggtcaa acgctcaaaa gcatgcctac ggcggacggt
     4741 ctcaaaccgt tgaaaaactt tgcttccttg caagaactgc tatcgctggg cggcgagcgc
     4801 cttctggcgg atttggtcag ggaaaacatg cgagtcaggg acatgcttaa cgaagtggcc
     4861 cccctgctca gggatgacgg cagctgcagc tctcttaact accagttgca gccggtaata
     4921 ggtgtgattt acgggcccac cggctgcggt aagtcgcagc tgctcaggaa cctgctttct
     4981 tcccagctga tctcccctac cccggaaacc gttttcttca tcgccccgca ggtagacatg
     5041 atccccccat ctgaactcaa agcgtgggaa atgcaaatct gtgagggtaa ctacgcccct
     5101 gggccggatg gaaccattat accgcagtct ggcaccctcc gcccgcgctt tgtaaaaatg
     5161 gcctatgacg atctcatcct ggaacacaac tatgacgtta gtgatcccag aaatatcttc
     5221 gcccaggccg ccgcccgtgg gcccattgcc atcattatgg acgaatgcat ggaaaatctt
     5281 ggaggtcaca agggcgtctc caagttcttc cacgcatttc cttctaagct acatgacaaa
     5341 tttcccaagt gcaccggata cactgtgctg gtggttctgc acaacatgaa tccccggagg
     5401 gatatggctg ggaacatagc caacctaaaa atacagtcca agatgcatct catatcccca
     5461 cgtatgcacc catcccagct taaccgcttt gtaaacactt acaccaaggg cctgcccctg
     5521 gcaatcagct tgctactgaa agacattttt aggcaccacg cccagcgctc ctgctacgac
     5581 tggatcatct acaacaccac cccgcagcat gaagctctgc agtggtgcta cctccacccc
     5641 agagacgggc ttatgcccat gtatctgaac atccagagtc acctttacca cgtcctggaa
     5701 aaaatacaca ggaccctcaa cgaccgagac cgctggtccc gggcctaccg cgcgcgcaaa
     5761 acccctaaat aaagacagca agacacttgc ttgatccaaa tccaaacaga gtctggtttt
     5821 ttatttatgt tttaaaccgc attgggaggg gaggaagcct tcagggcaga aacctgctgg
     5881 cgcagatcca acagctgctg agaaacgaca ttaagttccc gggtcaaaga atccaattgt
     5941 gccaaaagag ccgtcaactt gtcatcgcgg gcggatgaac gggaagctgc actgcttgca
     6001 agcgggctca ggaaagcaaa gtcagtcaca atcccgcggg cggtggctgc agcggctgaa
     6061 gcggcggcgg aggctgcagt ctccaacggc gttccagaca cggtctcgta ggtcaaggta
     6121 gtagagtttg cgggcaggac ggggcgacca tcaatgctgg agcccatcac attctgacgc
     6181 accccggccc atgggggcat gcgcgttgtc aaatatgagc tcacaatgct tccatcaaac
     6241 gagttggtgc tcatggcggc ggcggctgct gcaaaacaga tacaaaacta cataagaccc
     6301 ccaccttata tattctttcc cacccttaac cacgcccaga tcctctagca gtgataaacg
     6361 tctaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     6421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     6481 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     6541 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     6601 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     6661 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     6721 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     6781 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     6841 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     6901 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag agatctcgag
     6961 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccgcc cctctccctc
     7021 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     7081 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     7141 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     7201 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     7261 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     7321 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     7381 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     7441 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtaca catgctttac
     7501 atgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     7561 cctttgaaaa acacgatgat aatatggcca caaccatggt gagcaagggc gaggagctgt
     7621 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     7681 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     7741 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     7801 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     7861 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     7921 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     7981 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     8041 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     8101 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     8161 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     8221 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     8281 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgctcgagc ctaagcttct
     8341 agataagata tccgatccac cggatctaga taactgatca taatcagcca taccacattt
     8401 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     8461 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     8521 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     8581 tccaaactca tcaatgtatc ttaacgcgat caagctagct tgctagactc gactgactat
     8641 aataataaaa cgccaacttt gacccggaac gcggaaaaca cctgagaaaa acacctgggc
     8701 gagtctccac gtaaacggtc aaagtccccg cggccctaga caaatattac gcgctatgag
     8761 taacacaaaa ttattcagat ttcacttcct cttattcagt tttcccgcga aaatggccaa
     8821 atcttactcg gttacgccca aatttactac aacatccgcc taaaaccgcg cgaaaattgt
     8881 cacttcctgt gtacaccggc gcacaccaaa aacgtcactt ttgccacatc cgtcgcttac
     8941 atgtgttccg ccacacttgc aacatcacac t
//
