LOCUS       Exported                8276 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 8276)
  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 8276)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 14, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8276
                     /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..3206
                     /label=TCR-E
     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    3213..3789
                     /label=IRES
     primer_bind     complement(3382..3399)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3609..3628
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     misc_feature    3790..4509
                     /label=mAmetrine 1.1
                     /label=mAmetrine\1.1
                     /note="/vntifkey=21"
     primer_bind     complement(3835..3856)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4096..4115)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4443..4463
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4671..5185
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5347..5363
                     /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(5371..5401)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5416..5437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5554..5571)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5725..6313)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5805..5824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6484..7344)
                     /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     7107..7126
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7345..7449)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7517..7535
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7573..7595)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7695..7714
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7908..7930
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8058..8077
                     /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 tcatgaacaa ttccccagct ttagtgactg
     1441 tgatgctgtt tatacttggg aggacccatg gagactcagt gatccagatg caaggtcaag
     1501 tgaccctttc agaaaacgac ttcttattta taaactgcac ttattcaacc acagggtacc
     1561 cgactctttt ctggtatgtc caatattctg gagaaggtcc acagctcctt ctgcaagtca
     1621 caactgccaa caacaaggga agcagcagag gttttgaagc tacatatgat aaaggaacca
     1681 cttccttcca cttgcagaaa acctcagtgc aagagataga ttcggctgtg tactactgtg
     1741 ctatgagaga cacaaatgct tacaaagtca tctttggaaa agggacacat cttcatgttc
     1801 tccctaacat 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 tgaacaagtg ggttttctgc tgggtaaccc tttgtctcct tactgtagag accacacatg
     2341 gtgatggtgg catcattact cagacaccca aattcctgat tggtcaggaa gggcaaaaac
     2401 tgaccttgaa atgtcaacag aatttcaatc atgatacaat gtactggtac cgacaggatt
     2461 cagggaaagg attgagactg atctactatt caataactga aaacgatctt caaaaaggcg
     2521 atctatctga aggctatgat gcgtctcgag agaagaagtc atctttttct ctcactgtga
     2581 catctgccca gaagaacgag atggccgttt ttctctgtgc cagcagtata ggcaataaca
     2641 accaggctcc gctttttgga gaggggactc gactttctgt tctagaggat ctgagaaatg
     2701 tgactccacc caaggtctcc ttgtttgagc catcaaaagc agagattgca aacaaacaaa
     2761 aggctaccct cgtgtgcttg gccaggggct tcttccctga ccacgtggag ctgagctggt
     2821 gggtgaatgg caaggaggtc cacagtgggg tcagcacgga ccctcaggcc tacaaggaga
     2881 gcaattatag ctactgcctg agcagccgcc tgagggtctc tgctaccttc tggcacaatc
     2941 ctcgaaacca cttccgctgc caagtgcagt tccatgggct ttcagaggag gacaagtggc
     3001 cagagggctc acccaaacct gtcacacaga acatcagtgc agaggcctgg ggccgagcag
     3061 actgtggaat cacttcagca tcctatcatc agggggttct gtctgcaacc atcctctatg
     3121 agatcctact ggggaaggcc accctatatg ctgtgctggt cagtggcctg gtgctgatgg
     3181 ccatggtcaa gaaaaaaaat tcctgactcg agcgggatca attccgcccc ccccctaacg
     3241 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
     3301 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
     3361 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     3421 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     3481 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     3541 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     3601 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     3661 cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga
     3721 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac
     3781 gataatacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3841 gagctggacg gcgacgtaaa cggccacagg ttcagcgtgc gcggcgaggg cgagggcgat
     3901 gccaccaacg gcaagctgac cctgaagttc atctgcacct ccggcgagct gcccgtgccc
     3961 tggcccaccc tcgtgaccac cctgtcttac ggcgtgcagt gcttcgcccg ctaccccgac
     4021 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4081 accatctcct tcaaggacga cggcagctac aggacccgcg ccgaggtgaa gttcgagggc
     4141 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4201 ctggggcaca agctggagta caacatgaac gcgtgggacg cgtatatcac ggccgacaag
     4261 cagaagaacg gcatcaaagc gaacttcaag atcgagcaca acgtcaagga cggcggcgtg
     4321 cagctcgccg acgcgtacca gcagaacacc cccatcggcg acggctccgt gctgctgcct
     4381 gacaaccact acctgagctt ccagagcaag ctgttcaaag accccaacga gcagcgcgat
     4441 cacatggtcc tgctggagtt cgttaccgcc gccgggataa ctcccggcat ggacgagctg
     4501 tacaagtaaa gcggccgcga ctctagagtc gacctgcagg catgcaagct tcaggtagcc
     4561 ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa tttaaagcgc
     4621 tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     4681 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     4741 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     4801 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     4861 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     4921 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     4981 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     5041 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     5101 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     5161 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     5221 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     5281 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
     5341 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5401 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5461 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5521 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5581 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5641 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5701 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5761 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5821 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5881 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5941 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     6001 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     6061 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     6121 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6181 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6241 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6301 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6361 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6421 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6481 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6541 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6601 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6661 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6721 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6781 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6841 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6901 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6961 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     7021 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     7081 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     7141 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7201 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7261 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7321 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7381 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7441 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     7501 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     7561 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     7621 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     7681 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     7741 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     7801 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     7861 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     7921 ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     7981 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     8041 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     8101 ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac
     8161 aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata
     8221 gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa gggggg
//
