LOCUS       sequence-354437-        6502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a bicistronic IRES-mCherry containing retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6502)
  TITLE     Vignali lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6502)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMSCV-IRES-mCherry FP
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     387..404
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    521..542
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(557..562,563..580,581..587)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    595..611
                     /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)."
     LTR             773..1287
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     CDS             complement(1448..2158)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1726..1745)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1995..2013
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     misc_feature    2161..2710
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2320..2339)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     2549..2566
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2779..3195)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2795..2811)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2827..2849)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    3262..3603
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     LTR             3667..4183
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(4383..4402)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4530..4552)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4746..4765)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4865..4887
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4925..4943)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5011..5115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5116..5184,5185..5976)
                     /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     complement(5334..5353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6147..6502,1..233)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
       61 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
      121 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
      181 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
      241 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
      301 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
      361 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
      421 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      481 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
      541 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
      601 gcggataaca atttcacaat tctccccagc tctttctgcg ctgtccataa tgaactattt
      661 caggagtatt ggagtatgtg gattcaaaac agtggctaat tgagtcagga ttacttaata
      721 ttcgactcct accctcagaa tgttgttctc caacttcaag aaactgttac ccatgaaaga
      781 cccccgctga cgggtagtca atcactcaga ggagaccctc ccaaggaaca gcgagaccac
      841 aagtcggatg caactgcaag agggtttatt ggatacacgg gtacccgggc gacgcagtct
      901 atcggaggac tggcgcgccg agtgaggggt tgtgggctct tttattgagc tcggggagca
      961 gaagcgcgcg aacagaagcg agaagcgaac tgattggtta gttcaaataa ggcacagggt
     1021 catttcaggt ccttggggca ccctggaaac atctgatggt tctctagaaa ctgctgaggg
     1081 cgggaccgca tctggggacc atctgttctt ggccctgagc cggggcagga actgcttacc
     1141 acagatatcc tgtttggccc atattctgct gtctctctgt tcctaacctt gatctgaact
     1201 tctctattct cagttatgta ttttccatgc cttgcaaaat ggcgttactt aagctagctt
     1261 gccaaaccta caggtggggt ctttcattcc cccctttttc tggagactaa ataaaatctt
     1321 ttattttatc gatatcagcg ctttaaattt gcgcatgcta gctatagttc tagaggtacc
     1381 ggttgttaac gttagccggc tacctgaagc ttgcatgcct gcaggtcgac tctagagtcg
     1441 cggccgctta cttgtacagc tcgtccatgc cgccggtgga gtggcggccc tcggcgcgtt
     1501 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccaac ttgatgttga
     1561 cgttgtaggc gccgggcagc tgcacgggct tcttggcctt gtaggtggtc ttgacctcag
     1621 cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt gatctcgccc ttcagggcgc
     1681 cgtcctcggg gtacatccgc tcggaggagg cctcccagcc catcgtcttc ttctgcatta
     1741 cggggccgtc ggaggggaag ttggtgccgc gcagcttcac cttgtagatg aactcgccgt
     1801 cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc gtcctcgaag ttcatcacgc
     1861 gctcccactt gaagccctcg gggaaggaca gcttcaagta gtcggggatg tcggcggggt
     1921 gcttcacgta ggccttggag ccgtacatga actgagggga caggatgtcc caggcgaagg
     1981 gcagggggcc acccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     2041 cctcgccctc gccctcgatc tcgaactcgt ggccgttcac ggagccctcc atgtgcacct
     2101 tgaagcgcat gaactccttg atgatggcca tgttatcctc ctcgcccttg ctcaccatgg
     2161 tattatcgtg tttttcaaag gaaaaccacg tccccgtggt tcggggggcc tagacgtttt
     2221 tttaacctcg actaaacaca tgtaaagcat gtgcaccgag gccccagatc agatcccata
     2281 caatggggta ccttctgggc atccttcagc cccttgttga atacgcttga ggagagccat
     2341 ttgactcttt ccacaactat ccaactcaca acgtggcact ggggttgtgc cgcctttgca
     2401 ggtgtatctt atacacgtgg cttttggccg cagaggcacc tgtcgccagg tggggggttc
     2461 cgctgcctgc aaagggtcgc tacagacgtt gtttgtcttc aagaagcttc cagaggaact
     2521 gcttccttca cgacattcaa cagaccttgc attcctttgg cgagagggga aagaccccta
     2581 ggaatgctcg tcaagaagac agggccaggt ttccgggccc tcacattgcc aaaagacggc
     2641 aatatggtgg aaaataacat atagacaaac gcacaccggc cttattccaa gcggcttcgg
     2701 ccagtaacgt tagggggggg gcggaattga tcccgctcga gcaattggga tccgctagct
     2761 acgtaagatc tgaattccgg cgcctagaga aggagtgagg gctggataaa gggaggatcg
     2821 aggcggggtc gaacgaggag gttcaagggg gagagacggg gcggatggag gaagaggagg
     2881 cggaggctta gggtgtacaa agggcttgac ccagggaggg gggtcaaaag ccaaggcttc
     2941 ccaggtcacg atgtagggga cctggtctgg gtgtccatgc gggccaggtg aaaagacctt
     3001 gatcttaacc tgggtgatga ggtctcggtt aaaggtgccg tctcgcggcc atccgacgtt
     3061 aaaggttggc cattctgcag agcagaaggt aacccaacgt ctcttcttga catctaccga
     3121 ctggttgtga gcgatccgct cgacatcttt ccagtgacct aaggtcaaac ttaagggagt
     3181 ggtaacagtc tggccctaat tttcagacaa atacagaaac acagtcagac agagacaaca
     3241 cagaacgatg ctgcagcgct gcagcagaca agacgcgcgg cttcggttcc aaaccgaaag
     3301 caaaaattca gacggaggcg ggaactgttt taggttctcg tctcctacca gaaccacata
     3361 tcctgacggg gtcggattcc acatcgactc ccttcctcag gtcgggccac aaaaacggcc
     3421 cccaaagtcc ctgggacgtc tcccagggtt gcggccgggt gttcagaact cgtcagttcc
     3481 accacgggtc cgccagatac agagctagtt agctaactag tacagacgca ggcgcaaaca
     3541 ttagatgccg gcacaaacac gcacaaagac agagacagac acgaaacgac cgctggccag
     3601 cttacctccc ggcggggggg tcggtggtcc ctgggcaggg gtctccaaat ctcggtggaa
     3661 cctccaaatg aaagaccccc gaggtgggca gtcaatcaat ctgaggagac cctcccaagg
     3721 atcagcgagt ccacgattcg gatgcaaaca gcaagaggct ttattgggaa tacgggtacc
     3781 cgggcgacgc agtctatcgg aggactggcg cgccgagtga ggggttgtgg gctcttttat
     3841 tgagctcggg gagcagaagc gcgcgaacag aagcgagaag cgaactgatt ggttagttca
     3901 aataaggcac agggtcattt caggtccttg gggcaccctg gaaacatctg atggttctct
     3961 agaaactgct gagggcggga ccgcatctgg ggaccatctg ttcttggccc tgagccgggg
     4021 caggaactgc ttaccacaga tatcctgttt ggcccatatt ctgctgtctc tctgttccta
     4081 accttgatct gaacttctct attctcagtt atgtattttc catgccttgc aaaatggcgt
     4141 tacttaagct agcttgccaa acctacaggt ggggtctttc attcccccct ttttctggag
     4201 actaaataaa atcttttatt ttatctatgg ctcgtactct ataggcttca gctggtgata
     4261 ttgttgagtc aaaactagag cctggaccac tgatatcctg tctttaacaa attggactaa
     4321 tcgcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     4381 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     4441 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     4501 ctccttgcat gcaccattcc ttgcgccgtc gttttacaac gtcgtgactg ggaaaaccct
     4561 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4621 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     4681 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     4741 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4801 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4861 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4921 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4981 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5041 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5101 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5161 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5221 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5281 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5341 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5401 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5461 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5521 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5581 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5641 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5701 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5761 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5821 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5881 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5941 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6001 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6061 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6121 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6181 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6241 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6301 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6361 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6421 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6481 acacagccca gcttggagcg aa
//
