LOCUS       sequence-210495-        6361 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent Yes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6361)
  TITLE     Expression of wt and mt tagged-human genes and oncogenes
REFERENCE   2  (bases 1 to 6361)
  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: Yes-EGFP
FEATURES             Location/Qualifiers
     source          1..6361
                     /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      585..594
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2241..2250
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2247..2249,2250..2252,2253..2966)
                     /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(2247..2249,2250..2252,2253..2963)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2292..2313)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2553..2572)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2900..2921
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3089..3210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3126..3145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3180..3199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3217..3672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3354..3375)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3566..3585
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3699..3803
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3805..4162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3805..3996
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3828..3848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4013..4148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4197..4991
                     /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             4197..4991
                     /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(4251..4270)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4861..4880
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5179..5198)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5223..5270
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5599..6187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6088..6107
                     /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 attctgcagt cgacgccacc atgggctgca
      601 ttaaaagtaa agaaaacaaa agtccagcca ttaaatacag acctgaaaat actccagagc
      661 ctgtcagtac aagtgtgagc cattatggag cagaacccac tacagtgtca ccatgtccgt
      721 catcttcagc aaagggaaca gcagttaatt tcagcagtct ttccatgaca ccatttggag
      781 gatcctcagg ggtaacgcct tttggaggtg catcttcctc attttcagtg gtgccaagtt
      841 catatcctgc tggtttaaca ggtggtgtta ctatatttgt ggccttatat gattatgaag
      901 ctagaactac agaagacctt tcatttaaga agggtgaaag atttcaaata attaacaata
      961 cggaaggaga ttggtgggaa gcaagatcaa tcgctacagg aaagaatggt tatatcccga
     1021 gcaattatgt agcgcctgca gattccattc aggcagaaga atggtatttt ggcaaaatgg
     1081 ggagaaaaga tgctgaaaga ttacttttga atcctggaaa tcaacgaggt attttcttag
     1141 taagagagag tgaaacaact aaaggtgctt attccctttc tattcgtgat tgggatgaga
     1201 taaggggtga caatgtgaaa cactacaaaa ttaggaaact tgacaatggt ggatactata
     1261 tcacaaccag agcacaattt gatactctgc agaaattggt gaaacactac acagaacatg
     1321 ctgatggttt atgccacaag ttgacaactg tgtgtccaac tgtgaaacct cagactcaag
     1381 gtctagcaaa agatgcttgg gaaatccctc gagaatcttt gcgactagag gttaaactag
     1441 gacaaggatg tttcggcgaa gtgtggatgg gaacatggaa tggaaccacg aaagtagcaa
     1501 tcaaaacact aaaaccaggt acaatgatgc cagaagcttt ccttcaagaa gctcagataa
     1561 tgaaaaaatt aagacatgat aaacttgttc cactatatgc tgttgtttct gaagaaccaa
     1621 tttacattgt cactgaattt atgtcaaaag gaagcttatt agatttcctt aaggaaggag
     1681 atggaaagta tttgaagctt ccacagctgg ttgatatggc tgctcagatt gctgatggta
     1741 tggcatatat tgaaagaatg aactatattc accgagatct tcgggctgct aatattcttg
     1801 taggagaaaa tcttgtgtgc aaaatagcag actttggttt agcaaggtta attgaagaca
     1861 atgaatacac agcaagacaa ggtgcaaaat ttccaatcaa atggacagct cctgaagctg
     1921 cactgtatgg tcggtttaca ataaagtctg atgtctggtc atttggaatt ctgcaaacag
     1981 aactagtaac aaagggccga gtgccatatc caggtatggt gaaccgtgaa gtactagaac
     2041 aagtggagcg aggatacagg atgccgtgcc ctcagggctg tccagaatcc ctccatgaat
     2101 tgatgaatct gtgttggaag aaggaccctg atgaaagacc aacatttgaa tatattcagt
     2161 ccttcttgga agactacttc actgctacag agccacagta ccagccagga gaaaatttat
     2221 gggcccggga tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg
     2281 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2341 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2401 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2461 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2521 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2581 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2641 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     2701 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     2761 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2821 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     2881 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2941 tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac
     3001 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3061 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3121 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3181 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3241 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3301 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3361 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3421 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3481 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3541 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3601 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3661 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3721 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3781 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3841 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3901 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3961 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4021 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4081 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4141 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4201 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4261 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4321 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4381 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4441 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4501 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4561 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4621 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4681 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4741 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4801 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4861 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4921 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4981 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5041 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5101 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5161 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5221 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5281 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5341 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5401 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5461 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5521 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5581 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5641 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5701 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5761 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5821 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5881 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5941 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6001 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6061 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6121 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6181 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6241 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6301 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     6361 g
//
