LOCUS       Exported                8264 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 8264)
  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 8264)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 14, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8264
                     /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..3194
                     /label=TCR-B
     misc_feature    2214..2222
                     /label=linker
                     /note="/vntifkey=21"
     misc_feature    2217..2225
                     /label=linker
                     /note="/vntifkey=21"
     CDS             2223..2279
                     /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    3201..3777
                     /label=IRES
     primer_bind     complement(3370..3387)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3597..3616
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3778..4497
                     /label=mAmetrine 1.1
                     /label=mAmetrine\1.1
                     /note="/vntifkey=21"
     primer_bind     complement(3823..3844)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4084..4103)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4431..4451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4659..5173
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5335..5351
                     /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(5359..5389)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5404..5425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5542..5559)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5713..6301)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5793..5812)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6472..7332)
                     /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     7095..7114
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7333..7437)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7505..7523
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7561..7583)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7683..7702
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7896..7918
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8046..8065
                     /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 actaccacca
     1561 tgagggctgt gcagtggttc cgaaagaatt ccaggggcag cctcatcaat ctgttctact
     1621 tggcttcagg aacaaaggag aatgggaggc taaagtcagc atttgattct aaggagcgct
     1681 acagcaccct gcacatcagg gatgcccagc tggaggactc aggcacttac ttctgtgctg
     1741 ctggggcatc ttctggcagc tggcaactca tctttggatc tggaacccaa ctgacagtta
     1801 tgcctgacat ccagaaccca gaacctgctg tgtaccagtt aaaagatcct cggtctcagg
     1861 acagcaccct ctgcctgttc accgactttg actcccaaat caatgtgccg aaaaccatgg
     1921 aatctggaac gttcatcact gacaaaactg tgctggacat gaaagctatg gattccaaga
     1981 gcaatggggc cattgcctgg agcaaccaga caagcttcac ctgccaagat atcttcaaag
     2041 agaccaacgc cacctacccc agttcagacg ttccctgtga tgccacgttg actgagaaaa
     2101 gctttgaaac agatatgaac ctaaactttc aaaacctgtc agttatggga ctccgaatcc
     2161 tcctgctgaa agtagccgga tttaacctgc tcatgacgct gaggctgtgg tccggctccg
     2221 gagccacgaa cttctctctg ttaaagcaag caggagacgt ggaagaaaac cccggtccca
     2281 tgggctccag gctctttctg gtcttgagcc tcctgtgtac aaaacacatg gaggctgcag
     2341 tcacccaaag ccctagaaac aaggtgacag taacaggagg aaacgtgaca ttgagctgtc
     2401 gccagactaa tagccacaac tacatgtact ggtatcggca ggacactggg catgggctga
     2461 ggctgatcca ttactcatat ggtgctggca accttcaaat aggagatgtc cctgatgggt
     2521 acaaggccac cagaacaacg caagaagact tcttcctcct gctggaattg gcttctccct
     2581 ctcagacatc tttgtacttc tgtgccagca gcgcgggact ggggagctcc tatgaacagt
     2641 acttcggtcc cggcaccagg ctcacggttt tagaggatct gagaaatgtg actccaccca
     2701 aggtctcctt gtttgagcca tcaaaagcag agattgcaaa caaacaaaag gctaccctcg
     2761 tgtgcttggc caggggcttc ttccctgacc acgtggagct gagctggtgg gtgaatggca
     2821 aggaggtcca cagtggggtc agcacggacc ctcaggccta caaggagagc aattatagct
     2881 actgcctgag cagccgcctg agggtctctg ctaccttctg gcacaatcct cgaaaccact
     2941 tccgctgcca agtgcagttc catgggcttt cagaggagga caagtggcca gagggctcac
     3001 ccaaacctgt cacacagaac atcagtgcag aggcctgggg ccgagcagac tgtggaatca
     3061 cttcagcatc ctatcatcag ggggttctgt ctgcaaccat cctctatgag atcctactgg
     3121 ggaaggccac cctatatgct gtgctggtca gtggcctggt gctgatggcc atggtcaaga
     3181 aaaaaaattc ctgactcgag cgggatcaat tccgcccccc ccctaacgtt actggccgaa
     3241 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
     3301 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
     3361 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
     3421 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
     3481 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
     3541 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
     3601 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
     3661 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
     3721 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga taataccatg
     3781 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     3841 gacgtaaacg gccacaggtt cagcgtgcgc ggcgagggcg agggcgatgc caccaacggc
     3901 aagctgaccc tgaagttcat ctgcacctcc ggcgagctgc ccgtgccctg gcccaccctc
     3961 gtgaccaccc tgtcttacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag
     4021 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catctccttc
     4081 aaggacgacg gcagctacag gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     4141 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     4201 ctggagtaca acatgaacgc gtgggacgcg tatatcacgg ccgacaagca gaagaacggc
     4261 atcaaagcga acttcaagat cgagcacaac gtcaaggacg gcggcgtgca gctcgccgac
     4321 gcgtaccagc agaacacccc catcggcgac ggctccgtgc tgctgcctga caaccactac
     4381 ctgagcttcc agagcaagct gttcaaagac cccaacgagc agcgcgatca catggtcctg
     4441 ctggagttcg ttaccgccgc cgggataact cccggcatgg acgagctgta caagtaaagc
     4501 ggccgcgact ctagagtcga cctgcaggca tgcaagcttc aggtagccgg ctaacgttaa
     4561 caaccggtac ctctagaact atagctagca tgcgcaaatt taaagcgctg atatcgataa
     4621 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4681 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat
     4741 agagaagttc agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata
     4801 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg
     4861 tcccgccctc agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg
     4921 aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc
     4981 gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct
     5041 ccgatagact gcgtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc
     5101 gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc
     5161 gggggtcttt catgggtaac agtttcttga agttggagaa caacattctg agggtaggag
     5221 tcgaatatta agtaatcctg actcaattag ccactgtttt gaatccacat actccaatac
     5281 tcctgaaata gttcattatg gacagcgcag aaagagctgg ggagaattgt gaaattgtta
     5341 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5401 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5461 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5521 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5581 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5641 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5701 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5761 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5821 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5881 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5941 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     6001 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     6061 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     6121 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     6181 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     6241 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     6301 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6361 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6421 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6481 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6541 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6601 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6661 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6721 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6781 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6841 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6901 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6961 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     7021 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     7081 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     7141 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     7201 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     7261 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     7321 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7381 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7441 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7501 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     7561 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     7621 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     7681 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     7741 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     7801 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     7861 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     7921 ggcgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     7981 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     8041 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     8101 gccacgatgc gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg
     8161 gtccaggctc tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca
     8221 tagataaaat aaaagatttt atttagtctc cagaaaaagg gggg
//
