LOCUS       Exported                7858 bp DNA     circular SYN 23-JUN-2021
DEFINITION  a bicistronic IRES-mCherry containing retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    pMSCV-IRES-mCherry FP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7858)
  TITLE     Vignali lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jun 23, 2021 from SnapGene Viewer 5.3.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
FEATURES             Location/Qualifiers
     source          1..7858
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     26..48
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     primer_bind     176..195
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
                     /note="color: black; sequence: GGTGATGTCGGCGATATAGG"
     LTR             395..911
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
                     /note="color: #ffe4c4"
     misc_feature    975..1316
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
                     /note="color: #ffe4c4"
     CDS             1383..1799
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
                     /note="color: #993366; direction: RIGHT"
     primer_bind     1729..1751
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
                     /note="color: black; sequence: CCCTTGAACCTCCTCGTTCGACC"
     primer_bind     1767..1783
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
                     /note="color: black; sequence: CTTTATCCAGCCCTCAC"
     misc_feature    1831..3087
                     /label=Covid-19 nucleocapsid phosphoprotein
                     /note="color: #ffcc99"
     CDS             3097..3123
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     CDS             3124..3150
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     CDS             3151..3177
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     misc_feature    3213..3760
                     /label=IRES
                     /note="color: #a6acb3"
     primer_bind     complement(3296..3315)
                     /label=Primer R (Alexjandro has)
                     /note="color: black; sequence: cctcacattgccaaaagacg; added:
                     2020-09-23"
     primer_bind     complement(3368..3385)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
                     /note="color: black; sequence: GCATTCCTTTGGCGAGAG"
     primer_bind     3595..3614
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
                     /note="color: black; sequence: TGGCTCTCCTCAAGCGTATT"
     CDS             3776..4486
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /note="color: #ff0000"
                     /translation="MVSMDEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3921..3939)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
                     /note="color: black; sequence: TTGGTCACCTTCAGCTTGG"
     primer_bind     4189..4208
                     /label=mCherry-F
                     /note="mCherry, forward primer"
                     /note="color: black; sequence: CCCCGTAATGCAGAAGAAGA"
     LTR             4647..5161
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
                     /note="color: #ffe4c4"
     protein_bind    5323..5339
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5347..5377)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5347 .. 5353 / #ffffff / -10
                        2: 5354 .. 5371 / #ffffff
                        3: 5372 .. 5377 / #ffffff / -35"
     protein_bind    5392..5413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     primer_bind     complement(5530..5547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     rep_origin      complement(5701..6289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     complement(5781..5800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     CDS             complement(6460..7320)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6460 .. 7251 / #ccffcc
                        2: 7252 .. 7320 / #ccffcc / signal sequence
                     Cleavage site after base 7251"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7083..7102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
                     /note="color: black; sequence: ATAATACCGCGCCACATAGC"
     promoter        complement(7321..7425)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     7493..7511
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
                     /note="color: black; sequence: AATAGGCGTATCACGAGGC"
     primer_bind     complement(7549..7571)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
                     /note="color: black; sequence: CCGGGAGCTGCATGTGTCAGAGG"
     primer_bind     7671..7690
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
                     /note="color: black; sequence: CGGCATCAGAGCAGATTGTA"
ORIGIN
        1 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggcgcaagga
       61 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
      121 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
      181 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
      241 gtccggcgta gaggcgatta gtccaatttg ttaaagacag gatatcagtg gtccaggctc
      301 tagttttgac tcaacaatat caccagctga agcctataga gtacgagcca tagataaaat
      361 aaaagatttt atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg
      421 gcaagctagc ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga
      481 gaagttcaga tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct
      541 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
      601 cgccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
      661 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
      721 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
      781 atagactgcg tcgcccgggt acccgtattc ccaataaagc ctcttgctgt ttgcatccga
      841 atcgtggact cgctgatcct tgggagggtc tcctcagatt gattgactgc ccacctcggg
      901 ggtctttcat ttggaggttc caccgagatt tggagacccc tgcccaggga ccaccgaccc
      961 ccccgccggg aggtaagctg gccagcggtc gtttcgtgtc tgtctctgtc tttgtgcgtg
     1021 tttgtgccgg catctaatgt ttgcgcctgc gtctgtacta gttagctaac tagctctgta
     1081 tctggcggac ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac
     1141 gtcccaggga ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga
     1201 atccgacccc gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc
     1261 ctccgtctga atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc
     1321 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt
     1381 agggccagac tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc
     1441 ggatcgctca caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg
     1501 cagaatggcc aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca
     1561 tcacccaggt taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc
     1621 cctacatcgt gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg
     1681 tacaccctaa gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc
     1741 ctcgttcgac cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg
     1801 gaattcagat cttacgtagc tagcggatcc atgtctgata atggacccca aaatcagcga
     1861 aatgcacccc gcattacgtt tggtggaccc tcagattcaa ctggcagtaa ccagaatgga
     1921 gaacgcagtg gggcgcgatc aaaacaacgt cggccccaag gtttacccaa taatactgcg
     1981 tcttggttca ccgctctcac tcaacatggc aaggaagacc ttaaattccc tcgaggacaa
     2041 ggcgttccaa ttaacaccaa tagcagtcca gatgaccaaa ttggctacta ccgaagagct
     2101 accagacgaa ttcgtggtgg tgacggtaaa atgaaagatc tcagtccaag atggtatttc
     2161 tactacctag gaactgggcc agaagctgga cttccctatg gtgctaacaa agacggcatc
     2221 atatgggttg caactgaggg agccttgaat acaccaaaag atcacattgg cacccgcaat
     2281 cctgctaaca atgctgcaat cgtgctacaa cttcctcaag gaacaacatt gccaaaaggc
     2341 ttctacgcag aagggagcag aggcggcagt caagcctctt ctcgttcctc atcacgtagt
     2401 cgcaacagtt caagaaattc aactccaggc agcagtaggg gaacttctcc tgctagaatg
     2461 gctggcaatg gcggtgatgc tgctcttgct ttgctgctgc ttgacagatt gaaccagctt
     2521 gagagcaaaa tgtctggtaa aggccaacaa caacaaggcc aaactgtcac taagaaatct
     2581 gctgctgagg cttctaagaa gcctcggcaa aaacgtactg ccactaaagc atacaatgta
     2641 acacaagctt tcggcagacg tggtccagaa caaacccaag gaaattttgg ggaccaggaa
     2701 ctaatcagac aaggaactga ttacaaacat tggccgcaaa ttgcacaatt tgcccccagc
     2761 gcttcagcgt tcttcggaat gtcgcgcatt ggcatggaag tcacaccttc gggaacgtgg
     2821 ttgacctaca caggtgccat caaattggat gacaaagatc caaatttcaa agatcaagtc
     2881 attttgctga ataagcatat tgacgcatac aaaacattcc caccaacaga gcctaaaaag
     2941 gacaaaaaga agaaggctga tgaaactcaa gccttaccgc agagacagaa gaaacagcaa
     3001 actgtgactc ttcttcctgc tgcagatttg gatgatttct ccaaacaatt gcaacaatcc
     3061 atgagcagtg ctgactcaac tcaggccggt agttcctacc catacgatgt tccagattac
     3121 gcttatccct acgacgtgcc tgattatgca tacccatatg atgtccccga ctatgcctaa
     3181 ggatcccaat tgctcgagcg ggatcaattc cgcccccccc ctaacgttac tggccgaagc
     3241 cgcttggaat aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct
     3301 tttggcaatg tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt
     3361 ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct
     3421 ctggaagctt cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc
     3481 ccacctggcg acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag
     3541 gcggcacaac cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc
     3601 tcctcaagcg tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga
     3661 tctgatctgg ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt
     3721 ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggt
     3781 gagcatggac gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
     3841 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     3901 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     3961 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     4021 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     4081 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     4141 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     4201 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     4261 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     4321 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     4381 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     4441 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagcgg ccgcgactct
     4501 agagtcgacc tgcaggcatg caagcttcag gtagccggct aacgttaaca accggtacct
     4561 ctagaactat agctagcatg cgcaaattta aagcgctgat atcgataaaa taaaagattt
     4621 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4681 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     4741 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     4801 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     4861 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     4921 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     4981 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     5041 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     5101 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     5161 tgggtaacag tttcttgaag ttggagaaca acattctgag ggtaggagtc gaatattaag
     5221 taatcctgac tcaattagcc actgttttga atccacatac tccaatactc ctgaaatagt
     5281 tcattatgga cagcgcagaa agagctgggg agaattgtga aattgttatc cgctcacaat
     5341 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5401 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5461 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5521 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5581 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     5641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     5701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5881 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6121 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     6481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     6541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     6601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     6661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     7441 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     7501 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     7561 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     7621 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     7681 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     7741 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     7801 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcga
//
