LOCUS       sequence-211330-        5219 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express TCR zeta-GFP fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5219)
  TITLE     TCR zeta-GFP fusions
REFERENCE   2  (bases 1 to 5219)
  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: pEGFPN1zetaYF5,6
FEATURES             Location/Qualifiers
     source          1..5219
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      591..600
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(597..599,600..602,603..1316)
                     /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(597..599,600..602,603..1313)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(642..663)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(903..922)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1250..1271
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1439..1560
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1476..1495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1530..1549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1567..2022)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1704..1725)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1916..1935
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2049..2153
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2155..2512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2155..2346
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2178..2198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2363..2498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2547..3341
                     /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             2547..3341
                     /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(2601..2620)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3211..3230
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3529..3548)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3573..3620
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3949..4537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4438..4457
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4712..5015
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5016..5219
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5170..5190
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ggtttagtga accgtcagat ccgctagcgc taccggactc agatctcgag ctcaagcttc
       61 gaattctgat gaagtggaaa gtgtctgttc tcgcctgcat cctccacgtg cggttcccag
      121 gagcagaggc acagagcttt ggtctgctgg atcccaaact ctgctacttg ctagatggaa
      181 tcctcttcat ctacggagtc atcatcacag ccctgtacct gagagcaaaa ttcagcagga
      241 gtgcagagac cgctgccaac ctgcaggacc ccaaccagct ctacaatgag ctcaatctag
      301 ggcgaagaga ggaatatgac gtcttggaga agaagcgggc tcgggatcca gagatgggag
      361 gcaaacagca gaggaggagg aacccccagg aaggcgtata caatgcactg cagaaagaca
      421 agatggcaga agcctacagt gagatcggca caaaaggcga gaggcggaga ggcaaggggc
      481 acgatggcct tttccagggt ctcagcactg ccaccaagga cacctttgat gccctgcata
      541 tgcagaccct ggcccctcgc acggtaccgc gggcccggga tccaccggtc gccaccatgg
      601 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
      661 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
      721 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
      781 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
      841 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
      901 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
      961 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1021 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1081 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1141 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1201 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1261 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     1321 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1381 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1441 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1501 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1561 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1621 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1681 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1741 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1801 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1861 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     1921 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     1981 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2161 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2221 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2281 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2341 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2401 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2461 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     2521 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2581 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2641 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2701 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     2761 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2821 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     2881 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     2941 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3001 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3061 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3121 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3181 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3241 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3301 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3361 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3421 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3481 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     3541 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3601 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3661 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3721 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3781 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3841 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3901 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3961 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4021 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4081 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4141 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4201 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4261 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4321 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4381 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4441 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4501 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4561 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4621 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     4681 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     4741 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     4801 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     4861 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     4921 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     4981 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     5041 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     5101 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     5161 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagct
//
