LOCUS       Exported                8267 bp ds-DNA     circular SYN 10-NOV-2022
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 8267)
  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 8267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 14, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8267
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..3197
                     /label=TCR-A
     misc_feature    2217..2225
                     /label=linker
                     /note="/vntifkey=21"
     misc_feature    2220..2228
                     /label=linker
                     /note="/vntifkey=21"
     CDS             2226..2282
                     /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    3204..3780
                     /label=IRES
     primer_bind     complement(3373..3390)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3600..3619
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3781..4500
                     /label=mAmetrine 1.1
                     /label=mAmetrine\1.1
                     /note="/vntifkey=21"
     primer_bind     complement(3826..3847)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4087..4106)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4434..4454
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4662..5176
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5338..5354
                     /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(5362..5392)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5407..5428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5545..5562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5716..6304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5796..5815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6475..7335)
                     /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     7098..7117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7336..7440)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7508..7526
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7564..7586)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7686..7705
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7899..7921
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8049..8068
                     /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 tgagcggaga taaggtgaaa caaagtccct
     1501 cagcgctgag tctccaagaa ggaaccaatt ctgctctgag atgcaatttt tctatcgccg
     1561 cgacaactgt gcagtggttc ctacagaatc ccaggggcag cctcatcaat cttttttacc
     1621 tggttccagg aacaaaggag aatgggaggt taaagtcagc attcgattct aaggagagct
     1681 acagcaccct gcacatcagg gatgcccagc tggaggactc aggcacttac ttctgtgctg
     1741 ctgggtctcc caactacagc aacaacagac ttactttggg gaagggaacc caggtggtgg
     1801 tgttaccaaa catccagaac ccagaacctg ctgtgtacca gttaaaagat cctcggtctc
     1861 aggacagcac cctctgcctg ttcaccgact ttgactccca aatcaatgtg ccgaaaacca
     1921 tggaatctgg aacgttcatc actgacaaaa ctgtgctgga catgaaagct atggattcca
     1981 agagcaatgg ggccattgcc tggagcaacc agacaagctt cacctgccaa gatatcttca
     2041 aagagaccaa cgccacctac cccagttcag acgttccctg tgatgccacg ttgactgaga
     2101 aaagctttga aacagatatg aacctaaact ttcaaaacct gtcagttatg ggactccgaa
     2161 tcctcctgct gaaagtagcc ggatttaacc tgctcatgac gctgaggctg tggtccggct
     2221 ccggagccac gaacttctct ctgttaaagc aagcaggaga cgtggaagaa aaccccggtc
     2281 ccatgggctg taggctccta agctgtgtgg ccttctgcct cttgggaata ggccctttgg
     2341 agacggctgt tttccagact ccaaactatc atgtcacaca ggtgggaaat gaagtgtctt
     2401 tcaattgtaa gcaaactctg ggccacgata ctatgtattg gtacaagcaa gactctaaga
     2461 aattgctgaa gattatgttt agctacaata ataagcaact cattgtaaac gaaacagttc
     2521 caaggcgctt ctcacctcag tcttcagata aagctcattt gaatcttcga atcaagtctg
     2581 tagagccgga ggactctgct gtgtatctct gtgccagcag cagggactgg ggggggctag
     2641 agtacttcgg tcccggcacc aggctcacgg ttttagagga tctgagaaat gtgactccac
     2701 ccaaggtctc cttgtttgag ccatcaaaag cagagattgc aaacaaacaa aaggctaccc
     2761 tcgtgtgctt ggccaggggc ttcttccctg accacgtgga gctgagctgg tgggtgaatg
     2821 gcaaggaggt ccacagtggg gtcagcacgg accctcaggc ctacaaggag agcaattata
     2881 gctactgcct gagcagccgc ctgagggtct ctgctacctt ctggcacaat cctcgaaacc
     2941 acttccgctg ccaagtgcag ttccatgggc tttcagagga ggacaagtgg ccagagggct
     3001 cacccaaacc tgtcacacag aacatcagtg cagaggcctg gggccgagca gactgtggaa
     3061 tcacttcagc atcctatcat cagggggttc tgtctgcaac catcctctat gagatcctac
     3121 tggggaaggc caccctatat gctgtgctgg tcagtggcct ggtgctgatg gccatggtca
     3181 agaaaaaaaa ttcctgactc gagcgggatc aattccgccc cccccctaac gttactggcc
     3241 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     3301 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     3361 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     3421 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     3481 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     3541 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     3601 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     3661 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     3721 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgataatacc
     3781 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3841 ggcgacgtaa acggccacag gttcagcgtg cgcggcgagg gcgagggcga tgccaccaac
     3901 ggcaagctga ccctgaagtt catctgcacc tccggcgagc tgcccgtgcc ctggcccacc
     3961 ctcgtgacca ccctgtctta cggcgtgcag tgcttcgccc gctaccccga ccacatgaag
     4021 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatctcc
     4081 ttcaaggacg acggcagcta caggacccgc gccgaggtga agttcgaggg cgacaccctg
     4141 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4201 aagctggagt acaacatgaa cgcgtgggac gcgtatatca cggccgacaa gcagaagaac
     4261 ggcatcaaag cgaacttcaa gatcgagcac aacgtcaagg acggcggcgt gcagctcgcc
     4321 gacgcgtacc agcagaacac ccccatcggc gacggctccg tgctgctgcc tgacaaccac
     4381 tacctgagct tccagagcaa gctgttcaaa gaccccaacg agcagcgcga tcacatggtc
     4441 ctgctggagt tcgttaccgc cgccgggata actcccggca tggacgagct gtacaagtaa
     4501 agcggccgcg actctagagt cgacctgcag gcatgcaagc ttcaggtagc cggctaacgt
     4561 taacaaccgg tacctctaga actatagcta gcatgcgcaa atttaaagcg ctgatatcga
     4621 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     4681 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag
     4741 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
     4801 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4861 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4921 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4981 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     5041 cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     5101 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     5161 agcgggggtc tttcatgggt aacagtttct tgaagttgga gaacaacatt ctgagggtag
     5221 gagtcgaata ttaagtaatc ctgactcaat tagccactgt tttgaatcca catactccaa
     5281 tactcctgaa atagttcatt atggacagcg cagaaagagc tggggagaat tgtgaaattg
     5341 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     5401 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     5461 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     5521 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     5641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     5701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     5761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     5821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     5941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6121 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     6481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     6541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     6601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     6661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     6721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6781 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7081 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7381 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7441 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     7501 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     7561 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     7621 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     7681 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     7741 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     7801 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     7861 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     7921 gacggcgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     7981 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     8041 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     8101 ccggccacga tgcgtccggc gtagaggcga ttagtccaat ttgttaaaga caggatatca
     8161 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag
     8221 ccatagataa aataaaagat tttatttagt ctccagaaaa agggggg
//
