LOCUS       Exported                8282 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 8282)
  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 8282)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 14, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8282
                     /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..3212
                     /label=TCR-G
     misc_feature    2229..2237
                     /label=linker
                     /note="/vntifkey=21"
     misc_feature    2232..2240
                     /label=linker
                     /note="/vntifkey=21"
     CDS             2238..2294
                     /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    3219..3795
                     /label=IRES
     primer_bind     complement(3388..3405)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3615..3634
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3796..4515
                     /label=mAmetrine 1.1
                     /label=mAmetrine\1.1
                     /note="/vntifkey=21"
     primer_bind     complement(3841..3862)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4102..4121)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4449..4469
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4677..5191
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5353..5369
                     /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(5377..5407)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5422..5443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5560..5577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5731..6319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5811..5830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6490..7350)
                     /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     7113..7132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7351..7455)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7523..7541
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7579..7601)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7701..7720
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7914..7936
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8064..8083
                     /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 tcatgatgcg tcctgtcacc tgctcagttc
     1441 ttgtgctcct cctaatgctc aggaggagca atggcgatgg agactccgtg acccagacag
     1501 aaggcctggt cactctcaca gaagggttgc ctgtgatgct gaactgcacc tatcagacta
     1561 tttactcaaa tcctttcctt ttctggtatg tgcaacatct caatgaatcc cctcggctac
     1621 tcctgaagag cttcacagac aacaagagga ccgagcacca agggttccac gccactctcc
     1681 ataagagcag cagctccttc catctgcaga agtcctcagc gcagctgtca gactctgccc
     1741 tgtactactg tgctctgagt gacgaggact atgcaaacaa gatgatcttt ggcttgggaa
     1801 ccattttgag agtcagacct cacatccaga acccagaacc tgctgtgtac cagttaaaag
     1861 atcctcggtc tcaggacagc accctctgcc tgttcaccga ctttgactcc caaatcaatg
     1921 tgccgaaaac catggaatct ggaacgttca tcactgacaa aactgtgctg gacatgaaag
     1981 ctatggattc caagagcaat ggggccattg cctggagcaa ccagacaagc ttcacctgcc
     2041 aagatatctt caaagagacc aacgccacct accccagttc agacgttccc tgtgatgcca
     2101 cgttgactga gaaaagcttt gaaacagata tgaacctaaa ctttcaaaac ctgtcagtta
     2161 tgggactccg aatcctcctg ctgaaagtag ccggatttaa cctgctcatg acgctgaggc
     2221 tgtggtccgg ctccggagcc acgaacttct ctctgttaaa gcaagcagga gacgtggaag
     2281 aaaaccccgg tcccatgagc tgcaggcttc tcctctatgt ttccctatgt cttgtggaaa
     2341 cagcactcat gaacactaaa attactcagt caccaagata tctaatcctg ggaagagcaa
     2401 ataagtcttt ggaatgtgag caacatctgg gacataatgc tatgtactgg tataaacaga
     2461 gcgctgagaa gccgccagag ctcatgtttc tctacaatct taaacagttg attcgaaatg
     2521 agacggtgcc cagtcgtttt atacctgaat gcccagacag ctccaagcta cttttacata
     2581 tatctgccgt ggatccagaa gactcagctg tctatttttg tgccagcagc caaggggggg
     2641 ctgatcaaga cacccagtac tttgggccag gcactcggct cctcgtgtta gaggatctga
     2701 gaaatgtgac tccacccaag gtctccttgt ttgagccatc aaaagcagag attgcaaaca
     2761 aacaaaaggc taccctcgtg tgcttggcca ggggcttctt ccctgaccac gtggagctga
     2821 gctggtgggt gaatggcaag gaggtccaca gtggggtcag cacggaccct caggcctaca
     2881 aggagagcaa ttatagctac tgcctgagca gccgcctgag ggtctctgct accttctggc
     2941 acaatcctcg aaaccacttc cgctgccaag tgcagttcca tgggctttca gaggaggaca
     3001 agtggccaga gggctcaccc aaacctgtca cacagaacat cagtgcagag gcctggggcc
     3061 gagcagactg tggaatcact tcagcatcct atcatcaggg ggttctgtct gcaaccatcc
     3121 tctatgagat cctactgggg aaggccaccc tatatgctgt gctggtcagt ggcctggtgc
     3181 tgatggccat ggtcaagaaa aaaaattcct gactcgagcg ggatcaattc cgcccccccc
     3241 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     3301 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     3361 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     3421 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc
     3481 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     3541 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     3601 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     3661 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     3721 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     3781 aaacacgata ataccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     3841 ctggtcgagc tggacggcga cgtaaacggc cacaggttca gcgtgcgcgg cgagggcgag
     3901 ggcgatgcca ccaacggcaa gctgaccctg aagttcatct gcacctccgg cgagctgccc
     3961 gtgccctggc ccaccctcgt gaccaccctg tcttacggcg tgcagtgctt cgcccgctac
     4021 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     4081 gagcgcacca tctccttcaa ggacgacggc agctacagga cccgcgccga ggtgaagttc
     4141 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     4201 aacatcctgg ggcacaagct ggagtacaac atgaacgcgt gggacgcgta tatcacggcc
     4261 gacaagcaga agaacggcat caaagcgaac ttcaagatcg agcacaacgt caaggacggc
     4321 ggcgtgcagc tcgccgacgc gtaccagcag aacaccccca tcggcgacgg ctccgtgctg
     4381 ctgcctgaca accactacct gagcttccag agcaagctgt tcaaagaccc caacgagcag
     4441 cgcgatcaca tggtcctgct ggagttcgtt accgccgccg ggataactcc cggcatggac
     4501 gagctgtaca agtaaagcgg ccgcgactct agagtcgacc tgcaggcatg caagcttcag
     4561 gtagccggct aacgttaaca accggtacct ctagaactat agctagcatg cgcaaattta
     4621 aagcgctgat atcgataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     4681 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     4741 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     4801 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4861 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     4921 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     4981 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     5041 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtgta tccaataaac
     5101 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     5161 gattgactac ccgtcagcgg gggtctttca tgggtaacag tttcttgaag ttggagaaca
     5221 acattctgag ggtaggagtc gaatattaag taatcctgac tcaattagcc actgttttga
     5281 atccacatac tccaatactc ctgaaatagt tcattatgga cagcgcagaa agagctgggg
     5341 agaattgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5401 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5461 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5521 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5581 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5641 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5701 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5761 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5821 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5881 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5941 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6001 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6061 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6121 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6181 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6241 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6301 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6361 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6421 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6481 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6541 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6601 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6661 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6721 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6781 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6841 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6901 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6961 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7021 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7081 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7141 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7201 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7261 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7321 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7381 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7441 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     7501 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     7561 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     7621 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     7681 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     7741 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     7801 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     7861 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     7921 acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     7981 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     8041 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     8101 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt
     8161 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     8221 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     8281 gg
//
