LOCUS       pMIAII-TCR-F        8255 bp DNA     circular SYN 05-APR-2023
DEFINITION  a bicistronic IRES-GFP containing retroviral vector, MCS is expanded
            from pMSCV-IRES-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8255)
  AUTHORS   Holst J, Szymczak-Workman AL, Vignali KM, Burton AR, Workman CJ, 
            Vignali DA
  TITLE     Generation of T-cell receptor retrogenic mice.
  JOURNAL   Nat Protoc. 2006;1(1):406-17.
  PUBMED    17406263
REFERENCE   2  (bases 1 to 8255)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 5, 2023 from SnapGene 6.2.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Protoc."; date: "2006"; volume: "1(1)"; pages: "406-17"
FEATURES             Location/Qualifiers
     source          1..8255
                     /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"
     misc_feature    581..922
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             989..1405
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1335..1357
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1373..1389
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     gene            1413..3185
                     /label=TCR-F
     misc_feature    2205..2213
                     /label=linker
                     /note="/vntifkey=21"
     misc_feature    2208..2216
                     /label=linker
                     /note="/vntifkey=21"
     CDS             2214..2270
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     misc_feature    3192..3768
                     /label=IRES
     primer_bind     complement(3361..3378)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3588..3607
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3769..4488
                     /label=mAmetrine 1.1
                     /label=mAmetrine\1.1
                     /note="/vntifkey=21"
     primer_bind     complement(3814..3835)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4075..4094)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4422..4442
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4650..5164
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5326..5342
                     /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(5350..5380)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5395..5416
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5533..5550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5704..6292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5784..5803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6463..7323)
                     /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     7086..7105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7324..7428)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7496..7514
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7552..7574)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7674..7693
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7887..7909
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8037..8056
                     /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 acgagttctg
      721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
      781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
      841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
      901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
      961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
     1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1381 agccctcact ccttctctag gcgccggaat tcatgatgca gaggaacctg ggagctgtgc
     1441 tggggattct gtgggtgcag atttgctggg tgagaggaga tcaggtggag cagagtcctt
     1501 cagccctgag cctccacgag ggaaccggtt ctgctctgag atgcaatttt acgaccacca
     1561 tgagggctgt gcagtggttc caacagaatt ccaggggcag cctcatcaat ctgttctact
     1621 tggcttcagg aacaaaggag aatgggaggt taaagtcaac attcaattct aaggagagct
     1681 acagcaccct gcacatcagg gatgcccagc tggaggactc aggcacttac ttctgtgctg
     1741 ctgaatttgc aaacaagatg atctttggct tgggaaccat tttgagagtc agacctcaca
     1801 tccagaaccc agaacctgct gtgtaccagt taaaagatcc tcggtctcag gacagcaccc
     1861 tctgcctgtt caccgacttt gactcccaaa tcaatgtgcc gaaaaccatg gaatctggaa
     1921 cgttcatcac tgacaaaact gtgctggaca tgaaagctat ggattccaag agcaatgggg
     1981 ccattgcctg gagcaaccag acaagcttca cctgccaaga tatcttcaaa gagaccaacg
     2041 ccacctaccc cagttcagac gttccctgtg atgccacgtt gactgagaaa agctttgaaa
     2101 cagatatgaa cctaaacttt caaaacctgt cagttatggg actccgaatc ctcctgctga
     2161 aagtagccgg atttaacctg ctcatgacgc tgaggctgtg gtccggctcc ggagccacga
     2221 acttctctct gttaaagcaa gcaggagacg tggaagaaaa ccccggtccc atgggcatcc
     2281 agaccctctg ttgtgtgatc ttttatgttc tgatagcaaa tcacacagat gctggagtta
     2341 cccagacacc cagacatgag gtggcagaga aaggacaaac aataatcctg aagtgtgagc
     2401 cagtttcagg ccacaatgac cttttctggt acagacagac caagatacag ggactagagt
     2461 tgctgagcta cttccgcagc aagtctctta tggaagatgg tggggctttc aaggatcgat
     2521 tcaaagctga gatgctaaat tcatccttct ccactctgaa gattcaacct acagaaccca
     2581 aggactcagc tgtgtatctg tgtgccagca gtttaggggg gtatgaacag tacttcggtc
     2641 ccggcaccag gctcacggtt ttagaggatc tgagaaatgt gactccaccc aaggtctcct
     2701 tgtttgagcc atcaaaagca gagattgcaa acaaacaaaa ggctaccctc gtgtgcttgg
     2761 ccaggggctt cttccctgac cacgtggagc tgagctggtg ggtgaatggc aaggaggtcc
     2821 acagtggggt cagcacggac cctcaggcct acaaggagag caattatagc tactgcctga
     2881 gcagccgcct gagggtctct gctaccttct ggcacaatcc tcgaaaccac ttccgctgcc
     2941 aagtgcagtt ccatgggctt tcggaggagg acaagtggcc agagggctca cccaaacctg
     3001 tcacacagaa catcagtgca gaggcctggg gccgagcaga ctgtggaatc acttcagcat
     3061 cctatcatca gggggttctg tctgcaacca tcctctatga gatcctactg gggaaggcca
     3121 ccctatatgc tgtgctggtc agtggcctgg tgctgatggc catggtcaag agaaaaaatt
     3181 cctgactcga gcgggatcaa ttccgccccc cccctaacgt tactggccga agccgcttgg
     3241 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     3301 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     3361 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     3421 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     3481 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     3541 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     3601 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     3661 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     3721 ccccgaacca cggggacgtg gttttccttt gaaaaacacg ataataccat ggtgagcaag
     3781 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     3841 ggccacaggt tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc
     3901 ctgaagttca tctgcacctc cggcgagctg cccgtgccct ggcccaccct cgtgaccacc
     3961 ctgtcttacg gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc
     4021 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatctcctt caaggacgac
     4081 ggcagctaca ggacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     4141 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     4201 aacatgaacg cgtgggacgc gtatatcacg gccgacaagc agaagaacgg catcaaagcg
     4261 aacttcaaga tcgagcacaa cgtcaaggac ggcggcgtgc agctcgccga cgcgtaccag
     4321 cagaacaccc ccatcggcga cggctccgtg ctgctgcctg acaaccacta cctgagcttc
     4381 cagagcaagc tgttcaaaga ccccaacgag cagcgcgatc acatggtcct gctggagttc
     4441 gttaccgccg ccgggataac tcccggcatg gacgagctgt acaagtaaag cggccgcgac
     4501 tctagagtcg acctgcaggc atgcaagctt caggtagccg gctaacgtta acaaccggta
     4561 cctctagaac tatagctagc atgcgcaaat ttaaagcgct gatatcgata aaataaaaga
     4621 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     4681 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     4741 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     4801 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     4861 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     4921 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     4981 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     5041 tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     5101 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     5161 tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga gtcgaatatt
     5221 aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata ctcctgaaat
     5281 agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt atccgctcac
     5341 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5401 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5461 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7441 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     7501 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     7561 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     7621 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     7681 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     7741 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     7801 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     7861 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg
     7921 aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata
     7981 cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg
     8041 atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg
     8101 cgtccggcgt agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct
     8161 ctagttttga ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa
     8221 taaaagattt tatttagtct ccagaaaaag ggggg
//
