LOCUS       sequence-212997-        6113 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6113)
  TITLE     Johannes Schmid plasmids
REFERENCE   2  (bases 1 to 6113)
  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: TNFR2-YFP
FEATURES             Location/Qualifiers
     source          1..6113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2111..2120
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2117..2119,2120..2122,2123..2836)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2162..2183)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2423..2442)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2770..2791
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2959..3080
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2996..3015)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3050..3069
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3087..3542)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3224..3245)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3436..3455
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3569..3673
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3675..4032
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3675..3866
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3698..3718)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3883..4018
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4067..4861
                     /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             4067..4861
                     /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(4121..4140)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4731..4750
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5049..5068)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5093..5140
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5469..6057
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5958..5977
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggactcagat ctcgagacca ccatggcgcc cgtcgccgtc tgggccgcgc tggccgtcgg
      721 actggagctc tgggctgcgg cgcacgcctt gcccgcccag gtggcattta caccctacgc
      781 cccggagccc gggagcacat gccggctcag agaatactat gaccaaacag ctcagatgtg
      841 ctgcagcaaa tgctcgccgg gccaacatgc aaaagtcttc tgtaccaaga cctcggacac
      901 cgtgtgtgac tcctgtgagg acagcacata cacccagctc tggaactggg ttcccgagtg
      961 cttgagctgt ggctcccgct gtagctctga ccaggtggaa actcaagcct gcactcggga
     1021 acagaaccgc atctgcacct gcaggcccgg ctggtactgc gcgctgagca agcaggaggg
     1081 gtgccggctg tgcgcgccgc tgcgcaagtg ccgcccgggc ttcggcgtgg ccagaccagg
     1141 aactgaaaca tcagacgtgg tgtgcaagcc ctgtgccccg gggacgttct ccaacacgac
     1201 ttcacccacg gatatttgca ggccccacca gatctgtaac gtggtggcca tccctgggaa
     1261 tgcaagcatg gatgcagtct gcacgtccac gtcccccacc cggagtatgg ccccaggggc
     1321 agtacactta ccccagccag tgtccacacg atcccaacac acgcagccaa ctccagaacc
     1381 cagcactgct ccaagcacct ccttcctgct cccaatgggc cccagccccc cagctgaagg
     1441 gagcactggc gacttcgctc ttccagttgg actgattgtg ggtgtgacag ccttgggtct
     1501 actaataata ggagtggtga actgtgtcat catgacccag gtgaaaaaga agcccttgtg
     1561 cctgcagaga gaagccaagg tgcctcactt gcctgccggt aaggcccggg gtacacaggg
     1621 ccccgagcag cagcacctac tgatcacagc gccgagctcc agcagcagct ccctggagag
     1681 ctcggccagt gcgttggaca gaagggcgcc cactcggaac cagccacagg caccaggcgt
     1741 ggaggccagt ggggccgggg aggcccgggc cagcaccggg agctcagatt cttcccctgg
     1801 tggccatggg acccaggtca atgtcacctg catcgtgaac gtctgtagca gctctgacca
     1861 cagctcacag tgctcctccc aagccagctc cacaatggga gacacagatt ccagcccctc
     1921 ggagtccccg aaggacgagc aggtcccctt ctccaaggag gaatgtgcct ttcggtcaca
     1981 gctggagacg ccagagaccc tgctggggag caccgaagag aagcccctgc cccttggagt
     2041 gcctgatgct gggatgaagc ccagtcgaat tctgcagtcg acggtaccgc gggcccggga
     2101 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2161 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     2221 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     2281 cgtgccctgg cccaccctcg tgaccacctt cggctacggc ctgcagtgct tcgcccgcta
     2341 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     2401 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     2461 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     2521 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     2581 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     2641 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2701 gctgcccgac aaccactacc tgagctacca gtccgccctg agcaaagacc ccaacgagaa
     2761 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2821 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     2881 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2941 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3001 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3061 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     3121 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     3181 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     3241 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     3301 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     3361 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     3421 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     3481 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     3541 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3601 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3661 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3781 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3841 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3901 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3961 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4021 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     4081 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4141 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4201 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     4261 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4321 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4381 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4441 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4501 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4561 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     4621 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4681 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4741 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4801 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4861 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4921 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4981 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     5041 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     5101 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     5161 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     5221 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     5281 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     5341 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5401 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5461 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5521 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5581 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5641 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5701 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5761 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5821 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5881 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5941 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6001 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6061 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tca
//
