LOCUS       sequence-228474-        8065 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector containing TCR sequences.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8065)
  AUTHORS   Wang R, Pscheid R, Ghumra A, Kan LYL, Cochrane S, Fairclough L, 
            Alcocer MJC
  TITLE     Towards a surrogate system to express human lipid binding TCRs.
  JOURNAL   Biotechnol Lett. 2019 Oct;41(10):1095-1104. doi: 
            10.1007/s10529-019-02713-2. Epub 2019 Jul 26.
  PUBMED    31346817
REFERENCE   2  (bases 1 to 8065)
  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: 
            "10.1007/s10529-019-02713-2"; journalName: "Biotechnol Lett"; date: 
            "2019-10"; volume: "41"; issue: "10"; pages: "1095-1104"
COMMENT     Sequence Label: pMJA219= pcDNA3.1V5-completeTRABV-CMVp
FEATURES             Location/Qualifiers
     source          1..8065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     552..571
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    570..625
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     671..690
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     promoter        complement(1892..2095)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1921..1941)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2096..2399
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             3563..3604
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3614..3631
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3654..3671)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3660..3884
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3930..4358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4017..4036)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4227..4248
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4367..4387)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4372..4701
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4552..4687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4614..4633
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4768..5562
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4822..4841)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5432..5451
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5738..5859
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5775..5794)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5829..5848
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5908..5924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5908..5924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5921..5943)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5932..5948
                     /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(5956..5962,5963..5980,5981..5986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6001..6022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6139..6156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6310..6898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6390..6409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7069..7860,7861..7929))
                     /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     7692..7711
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7930..8034)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagtaagaga agagggacag ctatgactgg gagtagtcag gagaggagga
      301 aaaatctggc tagtaaaaca tgtaaggaaa attttaggga tgttaaagaa aaaaataaca
      361 caaaacaaaa tataaaaaaa atctaacctc aagtcaaggc ttttctatgg aataaggaat
      421 ggacagcagg gggctgtttc atatactgat gacctcttta tagccacctt tgttcatggc
      481 agccagcata tggcatatgt tgccaaactc taaaccaaat actcattctg atgttttaaa
      541 tgatttgccc tcccatatgt ccttccgagt gagagacaca aaaaattcca acacactatt
      601 gcaatgaaaa taaatttcct ttattagcca gaagtcagat gctcaagggg cttcatgatg
      661 tccccataat ttttggcaga gggaaaaaga tctcagtggt atttgtgagc cagggcattg
      721 gccacaccag ccaccacctt ctgataggca gcctgcacct gaggagtgaa ttgaattctc
      781 agcctctgct gtcctttctc ttgaccatgg ccatcagcac cagggcagac accagcacgg
      841 cgtacagggt ggccttgccc agcaggatct cgtacaggat ggtggcgctc agcacgccct
      901 gctggtagct ctcgcttgtg aagccacagt cggctctgcc ccaggcctca gcagacacga
      961 tctgggtcac gggcttggct ctgtcctggg tccactcgtc gttctcgctc aggccgtaaa
     1021 actgcacctg gcatctgaag tggtttctgg ggttctgcca gaaggtggcg gacactctca
     1081 gcctgctgga caggcagtat ctgctgtcgt tcagggcggg ctgctctttc agaggctggg
     1141 gatcggtgga cacgccgctg tgcacctctt tgccgttgac ccaccaagac agttccacgt
     1201 ggtcggggta gaagccggtg gccagacaca cgagggtggc tttctgggtg tgggagatct
     1261 cggcctcaga aggctcgaac acggccacct cgggtgggaa cacgttcttc agatcttcca
     1321 gcacggtcag tctgctgcca gcgccaaatg tcagcacgtt ggcgccaggt gttccgccca
     1381 gaggagagct gctggcacac aggtactggc tggtgtggct gggtctagcg ctttccaggg
     1441 tcagtgggaa gtgctcggtt ctgattctgg acacggtgct ctcgctgctc aggtcgccct
     1501 tctcggtgct gttcacgccg tagctgtagt ggatcaggtg cagttccatg ccggggtcct
     1561 gctgatacca gtacatcttg tcgtggccca tggtctggct gcattccagg gtgattttct
     1621 tgccggtgcc gatcacgagg tatctggggg tctggtagat gtcggcttcc atcaggccag
     1681 cgcccaggaa gtagaagccc acgtagcaca gcagtctgat ggtcatgaca gcatcccctt
     1741 gttcagggtc caggacttct gggggaacta tttggaggtt tggatctgta gtctcctaat
     1801 aggctctgct cgacaagctt atcgatgcgg ccgcgctagc acgcgtcagc tgactagagg
     1861 atccctagcg gatctgacgg ttcactaaac cagctctgct tatatagacc tcccaccgta
     1921 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     1981 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     2041 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     2101 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     2161 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     2221 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     2281 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     2341 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     2401 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactac
     2461 tcgagtatgt cgaggtggcg tgtacggtgg gaggcctata taagcagagc tcgtttagtg
     2521 aaccgtcaga tcgcctggag aattcaccgg tcatatgccg cgggggccca gatcctgtgt
     2581 gtgtgtgtgt gtgcttgtgc acatctgttc tctgattggc tgactgtact tcctgagagt
     2641 ccctgtttaa tgttacaaga aacagctgat gaaacaaggc agcagatttg aggcaggctt
     2701 ctccagaaca accatgaaga agcacctgac cacctttctc gtgatcctgt ggctgtactt
     2761 ctacagaggc aacggcaaga accaggtgga acagagcccc cagagcctga tcatcctgga
     2821 aggcaagaac tgcaccctgc agtgcaacta caccgtgtcc ccattcagca acctgagatg
     2881 gtacaagcag gacaccggca gaggccccgt gtccctgacc atcatgacct tcagcgagaa
     2941 caccaagagc aacggcagat acaccgccac cctggacgcc gacacaaagc agagcagcct
     3001 gcacatcacc gccagccagc tgagcgacag cgccagctac atctgcgtgg tgtccgacag
     3061 aggcagcacc ctgggcagac tgtacttcgg cagaggaacc cagctgaccg tgtggcccga
     3121 catccagaac cctgaccctg ccgtgtacca gctgagagac tccaagtcca gcgacaagag
     3181 cgtgtgcctg ttcaccgact tcgacagcca gaccaacgtg tcccagagca aggacagcga
     3241 cgtgtacatc accgacaaga ccgtgctgga catgagatcc atggacttca agtctaacag
     3301 cgccgtggcc tggtccaaca agagcgactt cgcctgcgcc aacgccttca acaacagcat
     3361 tatccccgag gacacattct tcccaagccc cgagagcagc tgcgacgtga agctggtgga
     3421 aaagagcttc gagacagaca ccaacctgaa cttccagaac ctgagcgtga tcggcttcag
     3481 aatcctgctg ctgaaggtgg ccggcttcaa cctgctgatg accctgagac tgtggtccag
     3541 ctgatctaga gggcccttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc
     3601 tacgcgtacc ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac
     3661 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3721 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3781 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3841 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3901 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3961 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4021 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4081 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4141 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4201 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4261 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4321 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4381 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4441 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4501 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4561 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4621 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4681 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4741 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4801 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4861 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4921 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4981 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     5041 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     5101 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5161 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5221 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5281 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5341 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5401 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5461 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5521 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5581 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5641 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5701 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5761 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5821 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5881 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     5941 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6001 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6061 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6121 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6181 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6241 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6301 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6361 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6421 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6481 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6541 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6601 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6661 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6721 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6781 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6841 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6901 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6961 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7021 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7081 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7141 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7201 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7261 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7321 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7381 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7441 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7501 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7561 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7621 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7681 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7741 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7801 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7861 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7921 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7981 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8041 tccccgaaaa gtgccacctg acgtc
//
