LOCUS       sequence-252503-        6610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A TCR Variable Region cloning expression vector using Mouse Constant
            Regions to reduce mis-pairing with endogenous TCR ref Biotechniques.
            2015 Mar 1;58(3):135-9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  AUTHORS   Coren LV, Jain S, Trivett MT, Ohlen C, Ott DE
  TITLE     Production of retroviral constructs for effective transfer and 
            expression of T-cell receptor genes using Golden Gate cloning.
  JOURNAL   Biotechniques. 2015 Mar 1;58(3):135-9. doi: 10.2144/000114265. 
            eCollection 2015.
  PUBMED    25757546
REFERENCE   2  (bases 1 to 6610)
  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: "10.2144/000114265";
            journalName: "Biotechniques"; date: "2015-03-1- 1"; volume: "58"; 
            issue: "3"; pages: "135-9"
COMMENT     Sequence Label: MSGV Mouse Acceptor
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..517
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     1357..1379
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1395..1411
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     1749..1765
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    2899..2951
                     /label=MCS
                     /note="multiple cloning site"
     LTR             2995..3509
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3681..3697
                     /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(3705..3711,3712..3729,3730..3735))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3750..3771
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3888..3905)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4059..4647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4139..4158)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4818..5609,5610..5678))
                     /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     5441..5460
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5679..5783)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5851..5869
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5907..5929)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6029..6048
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6242..6264
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6392..6411
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
       61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
      121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
      421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
      481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
      541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
      601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
      661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttcgg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttc gggggccgtt tttgtggccc
      781 gacctgagtc ctaaaatccc gatcgtttag gactctttgg tgcacccccc ttagaggagg
      841 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
      901 tgctttcggt ttgggaccga agccgcgccg cgcgtcttgt ctgctgcagc atcgttctgt
      961 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa tatgggcccg ggctagcctg
     1021 ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca
     1081 accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa
     1141 cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta
     1201 agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga
     1261 cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc
     1321 ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc
     1381 cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc atatggccat
     1441 atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct gacatgacaa
     1501 gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta gtccagcacg
     1561 aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg gtggtacctc
     1621 acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag aacctagaac
     1681 ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc aaagtagacg
     1741 gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg ggtggaccat
     1801 cctctagacc gcaacgttaa ttaagccacc aagtcttctt tttttgaaga cttcagaacc
     1861 ccgagcccgc cgtgtaccag ctgaaggacc ccagaagcca ggacagcacc ctgtgtctgt
     1921 tcaccgactt cgacagccag atcaacgtgc ccaagaccat ggaaagcggc accttcatca
     1981 ccgataagtg cgtgctggac atgaaggcca tggacagcaa gagcaacggc gccattgcct
     2041 ggtccaacca gaccagcttc acatgccagg acatcttcaa agagacaaac gccacctacc
     2101 cttccagcga cgtgccctgt gacgccaccc tgaccgagaa gtccttcgag acagacatga
     2161 atctgaattt ccagaacctg agcgtgatgg gcctgcggat cctgctgctg aaagtggccg
     2221 gcttcaacct gctgatgacc ctgcggctgt ggtccagccg ggccaagaga tctggcagcg
     2281 gcgccaccaa tttcagcctg ctgaaacagg ccggggacgt ggaagagaac cctaagtctt
     2341 ctttttgaag acttgatctg aggaacgtga ccccccccaa ggtgtccctg ttcgagccca
     2401 gcaaggccga gatcgccaac aagcagaaag ccaccctcgt gtgcctggcc agaggcttct
     2461 tccccgacca cgtggaactg tcctggtggg tcaacggcaa agaggtgcac agcggagtct
     2521 gcaccgaccc ccaggcttac aaagagagca actacagcta ctgcctgtcc agcagactgc
     2581 gggtgtccgc taccttctgg cacaaccccc ggaaccactt cagatgccag gtgcagttcc
     2641 acggcctgag cgaagaggac aagtggcccg agggcagccc caagcccgtg acccagaata
     2701 tcagcgccga ggcctggggc agagccgatt gtggcatcac cagcgccagc taccaccagg
     2761 gggtgctgag cgccaccatc ctgtacgaga tcctgctggg caaggccacc ctgtacgccg
     2821 tgctggtgtc cggactggtg ctgatggcca tggtcaagaa gaagaacagc tagcggccgc
     2881 tacgtagtcg acggatccct cgagctcaag cttcgaattc tgcagtcgac ggtaccgcgg
     2941 gcccgggatc cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     3001 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     3061 atacataact gagaatagag aagttcagat caaggttagg aacagagaga cagcagaata
     3121 tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc caagaacaga
     3181 tggtccccag atgcggtccc gccctcagca gtttctagag aaccatcaga tgtttccagg
     3241 gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc agttcgcttc
     3301 tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac aacccctcac
     3361 tcggcgcgcc agtcctccga tagactgcgt cgcccgggta cccgtgtatc caataaaccc
     3421 tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc ctctgagtga
     3481 ttgactaccc gtcagcgggg gtctttcatg ggtaacagtt tcttgaagtt ggagaacaac
     3541 attctgaggg taggagtcga atattaagta atcctgactc aattagccac tgttttgaat
     3601 ccacatactc caatactcct gaaatccatc gatggagttc attatggaca gcgcagaaag
     3661 agctggggag aattgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3721 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3781 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3841 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3901 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3961 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4021 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4081 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4141 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4201 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4261 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4321 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4381 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4441 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4501 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4561 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4621 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4681 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4741 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4801 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4861 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4921 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4981 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5041 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5101 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5161 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5221 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5281 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5341 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5401 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5461 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5521 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5581 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5641 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5701 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5761 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     5821 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     5881 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
     5941 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
     6001 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
     6061 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     6121 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     6181 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     6241 tcccagtcac gacgttgtaa aacgacggcg caaggaatgg tgcatgcaag gagatggcgc
     6301 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
     6361 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
     6421 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg cgattagtcc
     6481 aatttgttaa agacaggata tcagtggtcc aggctctagt tttgactcaa caatatcacc
     6541 agctgaagcc tatagagtac gagccataga taaaataaaa gattttattt agtctccaga
     6601 aaaagggggg
//
