LOCUS       sequence-164131-        5923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of TSAd-EGFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5923)
  TITLE     Spurkland Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5923)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pEGFP-TSAd-wt
FEATURES             Location/Qualifiers
     source          1..5923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1863..1872
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1869..1871,1872..1874,1875..2588)
                     /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(1869..1871,1872..1874,1875..2585)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1914..1935)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2175..2194)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2522..2543
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2711..2832
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2748..2767)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2802..2821
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2839..3294)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2976..2997)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3188..3207
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3321..3425
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3427..3784
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3427..3618
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3450..3470)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3635..3770
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3819..4613
                     /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             3819..4613
                     /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(3873..3892)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4483..4502
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4801..4820)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4845..4892
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5221..5809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5710..5729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttggt accgagctcg gatcccttgt gaggctgccc
      661 tcatggagtt ccccctggcc cagatatgtc cccaagggag tcacgaagcc cccatcccaa
      721 ccttcagcac cttccagatc acagacatga cccgcaggag ctgccagaac ctgggctaca
      781 ctgcggcatc tccccaggcc ccggaggctg cctccagcac agggaatgct gagagggcag
      841 aggaggtgcc tggagaagga agcctgttcc tgcaggccga gacccgggct tggttccaga
      901 agacccaggc ccactggctc ctgcagcacg gggcagcccc tgcctggttc catggcttca
      961 tcacccggag ggaggcagag aggctgctgg agcccaagcc tcaggggtgc tacttggtgc
     1021 ggttcagcga gagcgcggtg accttcgtgc tgacttacag gagccggact tgctgccgcc
     1081 acttcctgct ggcccagctc agggacgggc gccacgtggt gctgggcgag gacagcgccc
     1141 acgcgcggct gcaggacctg ctgctgcact acaccgcgca cccgctcagc ccctacgggg
     1201 agacgctcac cgagcccctc gcccgacaga ctcctgagcc tgcaggactt tccctgagga
     1261 ccgaagaatc aaactttgga agcaaaagcc aggacccaaa cccccagtac agcccaatca
     1321 tcaaacaggg gcaagcccca gtcccgatgc agaaagaggg ggccggggag aaggagccct
     1381 cccagctgct caggcccaag cctcccatcc ccgccaaacc tcagctgccc ccagaagtct
     1441 acacaatccc tgttccacga caccgcccgg ccccacgccc caagccctcc aatcctatct
     1501 acaatgagcc tgatgaaccc atagctttct atgccatggg ccggggcagc cctggggaag
     1561 cccccagcaa catctatgtg gaagtggaag atgagggcct acccgccacc cttgggcacc
     1621 ctgtcctacg gaagagctgg tccaggcctg tcccaggagg ccagaataca ggtggctccc
     1681 agctgcattc tgagaactct gtgattgggc aaggccctcc cctgccccac cagcccccac
     1741 ccgcctggag acacaccctc ccccacaatc tttctagaca ggtgcttcag gacagaggac
     1801 aggcatggct tccccttggg cctcctcact cgacggtacc gcgggcccgg gatccaccgg
     1861 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1921 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1981 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2041 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2101 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2161 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2221 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2281 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2341 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2401 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2461 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2521 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2581 acaagtaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2641 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2701 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2761 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2821 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2881 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2941 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3001 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3061 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3121 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3181 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3241 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3301 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3361 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3421 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3721 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3781 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4321 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4621 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4681 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4741 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4801 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4861 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4921 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4981 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5041 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5341 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5581 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5881 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
