LOCUS       sequence-231220-        6468 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a bicistronic IRES-DsRed 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 6468)
  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 6468)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Protoc."; date: "2006"; volume: "1(1)"; pages: "406-17"
COMMENT     Sequence Label: pMSCV-IRES-DsRed FP (pMIDsRed II)
FEATURES             Location/Qualifiers
     source          1..6468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    169..510
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             577..993
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     923..945
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     961..977
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1062..1611
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1206..1223)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1433..1452
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1614..2291
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(1744..1762)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(1794..1814)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     2012..2031
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2206..2229
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     LTR             2451..2965
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    3127..3143
                     /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(join(3151..3157,3158..3175,3176..3181))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3196..3217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3334..3351)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3505..4093)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3585..3604)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4264..5055,5056..5124))
                     /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     4887..4906
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5125..5229)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5297..5315
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5353..5375)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5475..5494
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5688..5710
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5838..5857
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(6057..6468,1..105)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gactcgctga tccttgggag
       61 ggtctcctca gattgattga ctgcccacct cgggggtctt tcatttggag gttccaccga
      121 gatttggaga cccctgccca gggaccaccg acccccccgc cgggaggtaa gctggccagc
      181 ggtcgtttcg tgtctgtctc tgtctttgtg cgtgtttgtg ccggcatcta atgtttgcgc
      241 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      301 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
      361 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
      421 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
      481 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
      541 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
      601 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
      661 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
      721 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
      781 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
      841 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
      901 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
      961 ctttatccag ccctcactcc ttctctaggc gccggaattc agatcttacg tagctagcgg
     1021 atcccaattg ctcgagcggg atcaattccg ccccccccct aacgttactg gccgaagccg
     1081 cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt
     1141 tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct
     1201 ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct
     1261 ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc
     1321 acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc
     1381 ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc
     1441 ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc
     1501 tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct
     1561 aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgataat accatggcct
     1621 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga
     1681 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     1741 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     1801 cccagttcca gtacggctcc aaggtgtacg tgaagcaccc cgccgacatc cccgactaca
     1861 agaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     1921 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg ctccttcatc tacaaggtga
     1981 agttcatcgg cgtgaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct
     2041 gggaggcctc caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccaca
     2101 aggccctgaa gctgaaggac ggcggccact acctggtgga gttcaagtcc atctacatgg
     2161 ccaagaagcc cgtgcagctg cccggctact actacgtgga ctccaagctg gacatcacct
     2221 cccacaacga ggactacacc atcgtggagc agtacgagcg cgccgagggc cgccaccacc
     2281 tgttcctgta gcggccgcga ctctagagtc gacctgcagg catgcaagct tcaggtagcc
     2341 ggctaacgtt aacaaccggt acctctagaa ctatagctag catgcgcaaa tttaaagcgc
     2401 tgatatcgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
     2461 ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac
     2521 ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg
     2581 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     2641 ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     2701 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     2761 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
     2821 cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt
     2881 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
     2941 ctacccgtca gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc
     3001 tgagggtagg agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac
     3061 atactccaat actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt
     3121 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3181 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3241 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3301 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3361 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3421 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3481 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3541 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3601 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3661 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3721 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3781 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3841 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3901 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3961 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4021 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4081 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4141 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4201 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4261 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4321 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4381 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4441 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4501 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4561 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4621 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4681 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4741 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4801 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4861 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4921 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4981 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5041 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5101 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5161 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5221 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     5281 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     5341 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     5401 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     5461 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
     5521 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     5581 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     5641 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     5701 ttgtaaaacg acggcgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg
     5761 gccacggggc ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga
     5821 gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg
     5881 ccggtgatgc cggccacgat gcgtccggcg tagaggcgat tagtccaatt tgttaaagac
     5941 aggatatcag tggtccaggc tctagttttg actcaacaat atcaccagct gaagcctata
     6001 gagtacgagc catagataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg
     6061 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     6121 aaaatacata actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga
     6181 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
     6241 agatggtccc cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc
     6301 agggtgcccc aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc
     6361 ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct
     6421 cactcggcgc gccagtcctc cgatagactg cgtcgcccgg gtacccgt
//
