LOCUS       sequence-356817-        6907 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse BTLA fused to mEGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6907)
  AUTHORS   Xu X, Hou B, Fulzele A, Masubuchi T, Zhao Y, Wu Z, Hu Y, Jiang Y, Ma
            Y, Wang H, Bennett EJ, Fu G, Hui E
  TITLE     PD-1 and BTLA regulate T cell signaling differentially and only 
            partially through SHP1 and SHP2.
  JOURNAL   J Cell Biol. 2020 Jun 1;219(6). pii: 151801. doi: 
            10.1083/jcb.201905085.
  PUBMED    32437509
REFERENCE   2  (bases 1 to 6907)
  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"; journalName: "J Cell 
            Biol."; date: "2020-06-1"; volume: "219(6). pii: 151801. doi"; 
            pages: " 10.1083/jcb.201905085"
COMMENT     Sequence Label: pMIG II-BTLA-mGFP
FEATURES             Location/Qualifiers
     source          1..6907
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(560..1351,1352..1420))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1183..1202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1421..1525)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1593..1611
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1649..1671)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1771..1790
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1984..2006
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2134..2153
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2353..2869
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2933..3274
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3341..3757
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3687..3709
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3725..3741
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      4767..4776
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4773..4775,4776..4778,4779..5492)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4818..4839)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5079..5098)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5426..5447
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5654..6168
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    6330..6346
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6354..6360,6361..6378,6379..6384))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6399..6420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6537..6554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6708..6907,1..389))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6788..6807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
       61 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
      121 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
      181 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
      241 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
      301 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
      361 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
      421 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
      481 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
      541 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
      601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
      781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
      841 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
      901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
      961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     1441 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     1501 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     1561 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     1621 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     1681 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     1741 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     1801 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
     1861 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     1921 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
     1981 tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc
     2041 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     2101 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
     2161 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt
     2221 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
     2281 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
     2341 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     2401 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     2461 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     2521 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     2581 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     2641 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     2701 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     2761 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg ctgatccttg
     2821 ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt ggaggttcca
     2881 ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag gtaagctggc
     2941 cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt
     3001 gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     3061 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
     3121 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
     3181 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     3241 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
     3301 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     3361 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     3421 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     3481 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     3541 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     3601 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     3661 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     3721 ctccctttat ccagccctca ctccttctct aggcgccgga attcgatctg gagctctcga
     3781 gaattctcac gcgtatgaag acagtgcctg ccatgcttgg gactcctcgg ttatttaggg
     3841 aattcttcat cctccatctg ggcctctgga gcatcctttg tgagaaagct actaagagga
     3901 atgatgaaga gtgtccagtg caacttacta ttacgaggaa ttccaaacag tctgccagga
     3961 caggagagtt atttaaaatt caatgtcctg tgaaatactg tgttcataga cctaatgtga
     4021 cttggtgtaa gcacaatgga acaatctgtg taccccttga ggttagccct cagctataca
     4081 ctagttggga agaaaatcaa tcagttccgg tttttgttct ccactttaaa ccaatacatc
     4141 tcagtgataa tgggtcgtat agctgttcta caaacttcaa ttctcaagtt attaatagcc
     4201 attcagtaac catccatgtg acagaaagga ctcaaaactc ttcagaacac ccactaataa
     4261 cagtatctga catcccagat gccaccaatg cctcaggacc atccaccatg gaagagaggc
     4321 caggcaggac ttggctgctt tacaccttgc ttcctttggg ggcattgctt ctgctccttg
     4381 cctgtgtctg cctgctctgc tttctgaaaa ggatccaagg gaaagaaaag aagccttctg
     4441 acttggcagg aagggacact aacctggttg atattccagc cagttccagg acaaatcacc
     4501 aagcactgcc atcaggaact ggaatttatg ataatgatcc ctggtctagc atgcaggatg
     4561 aatctgaatt gacaattagc ttgcaatcag agagaaacaa ccagggcatt gtttatgctt
     4621 ctttgaacca ttgtgttatt ggaaggaatc caagacagga aaacaacatg caggaggcac
     4681 ccacagaata tgcatccatt tgtgtgagaa gtacgcgtct gcaggatatc aagcttgcgg
     4741 taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     4801 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     4861 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     4921 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     4981 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     5041 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     5101 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     5161 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     5221 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     5281 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     5341 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     5401 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     5461 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga gtcgacctgc
     5521 aggcatgcaa gcttcaggta gccggctaac gttaacaacc ggtacctcta gaactatagc
     5581 tagcatgcgc aaatttaaag cgctgatatc gataaaataa aagattttat ttagtctcca
     5641 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     5701 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
     5761 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     5821 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
     5881 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     5941 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     6001 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     6061 ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg
     6121 gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcatgg gtaacagttt
     6181 cttgaagttg gagaacaaca ttctgagggt aggagtcgaa tattaagtaa tcctgactca
     6241 attagccact gttttgaatc cacatactcc aatactcctg aaatagttca ttatggacag
     6301 cgcagaaaga gctggggaga attgtgaaat tgttatccgc tcacaattcc acacaacata
     6361 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     6421 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     6481 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     6541 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     6601 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     6661 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     6721 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     6781 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     6841 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     6901 catagct
//
