LOCUS       sequence-378788-        7240 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human CBLB with FLAG tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7240)
  AUTHORS   Ge Y, Paisie TK, Chen S, Concannon P
  TITLE     UBASH3A Regulates the Synthesis and Dynamics of TCR-CD3 Complexes.
  JOURNAL   J Immunol. 2019 Dec 1;203(11):2827-2836. doi: 
            10.4049/jimmunol.1801338. Epub 2019 Oct 28.
  PUBMED    31659016
REFERENCE   2  (bases 1 to 7240)
  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: 
            "10.4049/jimmunol.1801338"; journalName: "J Immunol"; date: 
            "2019-12-1- 1"; volume: "203"; issue: "11"; pages: "2827-2836"
COMMENT     Sequence Label: FLAG-CBLB
FEATURES             Location/Qualifiers
     source          1..7240
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             74..868
                     /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             74..868
                     /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(128..147)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     738..757
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1056..1075)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1100..1147
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1476..2064
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1965..1984
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2239..2542
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2543..2746
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2697..2717
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        2792..2810
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2845..2854
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2854..2877
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5913..5932)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5914..5932)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    6206..6327
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6243..6262)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6297..6316
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6334..6789)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6471..6492)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6683..6702
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6816..6920
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(6922..7240,1..39)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6922..7113
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6945..6965)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      join(7130..7240,1..25)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
       61 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
      121 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
      181 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
      241 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
      301 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
      361 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
      421 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
      481 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
      541 tgatctggac gaagaacatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
      601 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
      661 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
      721 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagaacttg gcggcgaatg
      781 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
      841 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
      901 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
      961 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     1021 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     1081 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     1141 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     1201 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     1261 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     1321 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1381 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1441 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1501 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1561 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1621 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1681 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1741 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1801 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1861 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1921 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1981 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     2041 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     2101 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     2161 cgtattaccg ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag
     2221 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2281 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2341 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2401 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     2461 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     2521 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
     2581 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     2641 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     2701 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctggtt tagtgaaccg
     2761 tcagatccgc tagcgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc
     2821 tggagctcca ccgcggtggc ggccgccacc atggattaca aggatgacga cgataagagc
     2881 ccgggcggat cccccgggct gcagatggca aactcaatga atggcagaaa ccctggtggt
     2941 cgaggaggaa atccccgaaa aggtcgaatt ttgggtatta ttgatgctat tcaggatgca
     3001 gttggacccc ctaagcaagc tgccgcagat cgcaggaccg tggagaagac ttggaagctc
     3061 atggacaaag tggtaagact gtgccaaaat cccaaacttc agttgaaaaa tagcccacca
     3121 tatatacttg atattttgcc tgatacatat cagcatttac gacttatatt gagtaaatat
     3181 gatgacaacc agaaacttgc ccaactcagt gagaatgagt actttaaaat ctacattgat
     3241 agccttatga aaaagtcaaa acgggcaata agactcttta aagaaggcaa ggagagaatg
     3301 tatgaagaac agtcacagga cagacgaaat ctcacaaaac tgtcccttat cttcagtcac
     3361 atgctggcag aaatcaaagc aatctttccc aatggtcaat tccagggaga taactttcgt
     3421 atcacaaaag cagatgctgc tgaattctgg agaaagtttt ttggagacaa aactatcgta
     3481 ccatggaaag tattcagaca gtgccttcat gaggtccacc agattagctc tggcctggaa
     3541 gcaatggctc taaaatcaac aattgattta acttgcaatg attacatttc agtttttgaa
     3601 tttgatattt ttaccaggct gtttcagcct tggggctcta ttttgcggaa ttggaatttc
     3661 ttagctgtga cacatccagg ttacatggca tttctcacat atgatgaagt taaagcacga
     3721 ctacagaaat atagcaccaa acccggaagc tatattttcc ggttaagttg cactcgattg
     3781 ggacagtggg ccattggcta tgtgactggg gatgggaata tcttacagac catacctcat
     3841 aacaagccct tatttcaagc cctgattgat ggcagcaggg aaggatttta tctttatcct
     3901 gatgggagga gttataatcc tgatttaact ggattatgtg aacctacacc tcatgaccat
     3961 ataaaagtta cacaggaaca atatgaatta tattgtgaaa tgggctccac ttttcagctc
     4021 tgtaagattt gtgcagagaa tgacaaagat gtcaagattg agccttgtgg gcatttgatg
     4081 tgcacctctt gccttacggc atggcaggag tcggatggtc agggctgccc tttctgtcgt
     4141 tgtgaaataa aaggaactga gcccataatc gtggatccct ttgatccaag agatgaaggc
     4201 tccaggtgtt gcagcatcat tgaccccttt ggcatgccga tgctcgactt ggacgacgat
     4261 gatgatcgtg aggagtcctt gatgatgaat cggttggcaa acgtccgaaa gtgcactgac
     4321 aggcagaact caccagtcac atcaccagga tcctctcccc ttgcccagag aagaaagcca
     4381 cagcctgacc cactccagat cccacatcta agcctgccac ccgtgcctcc tcgcctggat
     4441 ctaattcaga aaggcatagt tagatctccc tgtggcagcc caacgggttc accaaagtct
     4501 tctccttgca tggtgagaaa acaagataaa ccactcccag caccacctcc tcccttaaga
     4561 gatcctcctc caccgccacc tgaaagacct ccaccaatcc caccagacaa tagactgagt
     4621 agacacatcc atcatgtgga aagcgtgcct tccaaagacc cgccaatgcc tcttgaagca
     4681 tggtgccctc gggatgtgtt tgggactaat cagcttgtgg gatgtcgact cctaggggag
     4741 ggctctccaa aacctggaat cacagcgagt tcaaatgtca atggaaggca cagtagagtg
     4801 ggctctgacc cagtgcttat gcggaaacac agacgccatg atttgccttt agaaggagct
     4861 aaggtctttt ccaatggtca ccttggaagt gaagaatatg atgttcctcc ccggctttct
     4921 cctcctcctc cagttaccac cctcctccct agcataaagt gtactggtcc gttagcaaat
     4981 tctctttcag agaaaacaag agacccagta gaggaagatg atgatgaata caagattcct
     5041 tcatcccacc ctgtttccct gaattcacaa ccatctcatt gtcataatgt aaaacctcct
     5101 gttcggtctt gtgataatgg tcactgtatg ctgaatggaa cacatggtcc atcttcagag
     5161 aagaaatcaa acatccctga cttaagcata tatttaaagg gagatgtttt tgattcagcc
     5221 tctgatcccg tgccattacc acctgccagg cctccaactc gggacaatcc aaagcatggt
     5281 tcttcactca acaggacgcc ctctgattat gatcttctca tccctccatt aggtgaagat
     5341 gcttttgatg ccctccctcc atctctccca cctcccccac ctcctgcaag gcatagtctc
     5401 attgaacatt caaaacctcc tggctccagt agccggccat cctcaggaca ggatcttttt
     5461 cttcttcctt cagatccctt tgttgatcta gcaagtggcc aagttccttt gcctcccgct
     5521 agaaggttac caggtgaaaa tgtcaaaact aacagaacat cacaggacta tgatcagctt
     5581 ccttcatgtt cagatggttc acaggcacca gccagacccc ctaaaccacg accgcgcagg
     5641 actgcaccag aaattcacca cagaaaaccc catgggcctg aggcggcatt ggaaaatgtc
     5701 gatgcaaaaa ttgcaaaact catgggagag ggttatgcct ttgaagaggt gaagagagcc
     5761 ttagagatag cccagaataa tgtcgaagtt gcccggagca tcctccgaga atttgccttc
     5821 cctcctccag tatccccacg tctaaatcta tagctcgagg gggggcccgg taccttaatt
     5881 aattaaggta ccaggtaagt gtacccaatt cgccctatag tgagtcgtat tacaattcac
     5941 tcgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggagatcc aatttttaag
     6001 tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc acagtcccaa
     6061 ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac
     6121 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     6181 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     6241 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6301 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     6361 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     6421 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     6481 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6541 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     6601 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6661 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     6721 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6781 agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6841 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6901 aatattgaaa aaggaagaat cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt
     6961 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     7021 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     7081 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     7141 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     7201 agaggccgag gccgcctcgg cctctgagct attccagaag
//
