LOCUS       sequence-250226-        5232 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CD3 delta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5232)
  AUTHORS   Lorenz H, Hailey DW, Lippincott-Schwartz J
  TITLE     Fluorescence protease protection of GFP chimeras to reveal protein 
            topology and subcellular localization.
  JOURNAL   Nat Methods. 2006 Mar;3(3):205-10. doi: 10.1038/nmeth857.
  PUBMED    16489338
REFERENCE   2  (bases 1 to 5232)
  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"; doi: "10"; journalName: 
            "Nat Methods"; date: "2006-03"; volume: "3"; issue: "3"; pages: 
            "205-10"
COMMENT     Sequence Label: CD3delta-CFP
FEATURES             Location/Qualifiers
     source          1..5232
                     /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      1172..1181
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1178..1180,1181..1183,1184..1897)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLK
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1223..1244)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1484..1503)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2020..2141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2057..2076)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2111..2130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2148..2603)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2285..2306)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2497..2516
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2630..2734
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2736..3093
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2736..2927
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2759..2779)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2944..3079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3128..3922
                     /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             3128..3922
                     /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(3182..3201)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3792..3811
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4110..4129)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4154..4201
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4530..5118
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5019..5038
                     /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 caagcttcga attcaccatg gaacacagcg ggattctggc
      661 tagtctgata ctgattgctg ttctccccca agggagcccc ttcaagatac aagtgaccga
      721 atatgaggac aaagtatttg tgacctgcaa taccagcgtc atgcatctag atggaacggt
      781 ggaaggatgg tttgcaaaga ataaaacact caacttgggc aaaggcgttc tggacccacg
      841 agggatatat ctgtgtaatg ggacagagca gctggcaaag gtggtgtctt ctgtgcaagt
      901 ccattaccga atgtgccaga actgtgtgga gctagactcg ggcaccatgg ctggtgtcat
      961 cttcattgac ctcatcgcaa ctctgctcct ggctttgggc gtctactgct ttgcaggaca
     1021 tgagaccgga aggccttctg gggctgctga ggttcaagca ctgctgaaga atgagcagct
     1081 gtatcagcct cttcgagatc gtgaagatac ccagtacagc cgtcttggag ggaactggcc
     1141 ccggaacaag aaatctggat cctcaccggt cgccaccatg gtgagcaagg gcgaggagct
     1201 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaggtt
     1261 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     1321 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctgggg
     1381 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     1441 catgcccgaa ggctacgtcc aggagcgtac catcttcttc aaggacgacg gcaactacaa
     1501 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     1561 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacatcag
     1621 ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggccc acttcaagat
     1681 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     1741 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     1801 gagcaaagac cccaacgaga agcgcgatca catggtcctt aaggagttcg tgaccgccgc
     1861 cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact ctagatcata
     1921 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     1981 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     2041 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     2101 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     2161 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     2221 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     2281 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     2341 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     2401 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     2461 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     2521 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     2581 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     2641 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     2701 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     2761 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2821 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2881 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2941 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     3001 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     3061 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     3121 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     3181 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     3241 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     3301 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     3361 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     3421 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     3481 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     3541 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     3601 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     3661 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3721 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3781 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3841 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3901 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3961 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     4021 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     4081 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     4141 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     4201 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     4261 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     4321 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     4381 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4441 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4501 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4561 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4621 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4681 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4741 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4801 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4861 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4921 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4981 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5041 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5101 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5161 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5221 accgccatgc at
//
