LOCUS       sequence-210749-        5793 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive Active.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5793)
  TITLE     Expression of wt and mt tagged-human genes and oncogenes
REFERENCE   2  (bases 1 to 5793)
  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: EGFP-PKCZdel239
FEATURES             Location/Qualifiers
     source          1..5793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1673..1682
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1679..1681,1682..1684,1685..2398)
                     /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(1679..1681,1682..1684,1685..2395)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1724..1745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1985..2004)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2332..2353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2521..2642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2558..2577)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2612..2631
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2649..3104)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2786..2807)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2998..3017
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3131..3235
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3237..3594
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3237..3428
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3260..3280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3445..3580
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3629..4423
                     /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             3629..4423
                     /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(3683..3702)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4293..4312
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4611..4630)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4655..4702
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5031..5619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5520..5539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatggga atcaaaatct ctcaggggct
      601 tgggctgcag gactttgacc taatcagagt catcgggcgc gggagctacg ccaaggttct
      661 cctggtgcgg ttgaagaaga atgaccaaat ttacgccatg aaagtggtga agaaagagct
      721 ggtgcatgat gacgaggata ttgactgggt acagacagag aagcacgtgt ttgagcaggc
      781 atccagcaac cccttcctgg tcggattaca ctcctgcttc cagacgacaa gtcggttgtt
      841 cctggtcatt gagtacgtca acggcgggga cctgatgttc cacatgcaga ggcagaggaa
      901 gctccctgag gagcacgcca ggttctacgc ggccgagatc tgcatcgccc tcaacttcct
      961 gcacgagagg gggatcatct acagggacct gaagctggac aacgtcctcc tggatgcgga
     1021 cgggcacatc aagctcacag actacggcat gtgcaaggaa ggcctgggcc ctggtgacac
     1081 aacgagcact ttctgcggaa ccccgaatta catcgccccc gaaatcctgc ggggagagga
     1141 gtacgggttc agcgtggact ggtgggcgct gggagtcctc atgtttgaga tgatggccgg
     1201 gcgctccccg ttcgacatca tcaccgacaa cccggacatg aacacagagg actacctttt
     1261 ccaagtgatc ctggagaagc ccatccggat cccccggttc ctgtccgtca aagcctccca
     1321 tgttttaaaa ggatttttaa ataaggaccc caaagagagg ctcggctgcc ggccacagac
     1381 tggattttct gacatcaagt cccacgcgtt cttccgcagc atagactggg acttgctgga
     1441 gaagaagcag gcgctccctc cattccagcc acagatcaca gacgactacg gtctggacaa
     1501 ctttgacaca cagttcacca gcgagcccgt gcagctgacc ccagacgatg aggatgccat
     1561 aaagaggatc gaccagtcag agttcgaagg ctttgagtat atcaacccat tattgctgtc
     1621 caccgaggag tcggtgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat
     1681 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1741 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1801 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1861 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1921 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1981 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2041 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2101 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2161 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2221 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2281 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2341 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag
     2401 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta
     2461 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     2521 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2581 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2641 taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2701 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2761 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2821 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2881 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2941 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     3001 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     3061 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     3121 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3181 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     3241 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3301 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3361 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3421 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3481 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3541 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3601 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3661 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3721 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3781 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3841 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3901 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3961 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4021 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4081 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4141 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4201 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4261 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4321 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4381 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4441 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4501 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4561 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4621 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4681 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4741 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4801 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4861 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4921 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4981 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5041 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5101 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5161 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5221 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5281 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5341 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5401 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5461 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5521 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5581 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5641 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5701 gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat caattacggg
     5761 gtcattagtt catagcccat atatggagtt ccg
//
