LOCUS       Exported                7872 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  retroviral expression of Runx1Fl and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    Banshee Runx1FL mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  TITLE     Rothenberg lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7872)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7872
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     58..80
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     96..112
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1547..1573
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1604..2171
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1621..2173
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1765..1782)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1992..2011
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      2194..2203
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2200..2910
                     /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(2345..2363)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2613..2632
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             2959..3473
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(3642..3658)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3642..3658)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3655..3677)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3666..3682
                     /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(3690..3720)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3735..3756
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3873..3890)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4044..4632)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4124..4143)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4803..5663)
                     /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     5426..5445
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5664..5768)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5836..5854
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5892..5914)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6014..6033
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6227..6249
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6377..6396
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             6596..7112
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    7176..7517
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             7584..128
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
ORIGIN
        1 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
       61 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
      121 taggcgccgg aattagatct ccaccatggc ttcagacagc atttttgagt catttccttc
      181 atatccacag tgcttcatga gagatgccag cacgagccgc cgcttcacgc cgccttccac
      241 cgcgctgagc cccggcaaga tgagcgaggc gctgccgctg ggcgccccgg atggcggcgc
      301 cgccctggcc agcaagctga ggagcggcga ccgcagcatg gtggaggtac tagctgacca
      361 ccctggcgag ctagtgcgca ccgacagccc caacttcctc tgctccgtgc tacccactca
      421 ctggcgctgc aacaagaccc tgcccatcgc tttcaaggtg gtggcactgg gggacgtccc
      481 ggatggcact ctggtcaccg tcatggcagg caacgatgaa aactactcgg cagaactgag
      541 aaatgctacc gcggccatga agaaccaggt agcgagattc aacgacctca ggtttgtcgg
      601 gcggagcggt agaggcaaga gcttcactct gaccatcacc gtctttacaa atccgccaca
      661 agttgccacc taccatagag ccatcaaaat cacagtggac ggcccccgag aaccccgaag
      721 acatcggcag aaactagatg atcagaccaa gcccgggagt ttgtcctttt ccgagcggct
      781 cagtgaattg gagcagctgc ggcgcacggc catgagggtc agcccgcacc acccagcccc
      841 cacgcccaac cctcgggcct ccttgaacca ctccactgcc tttaaccctc agcctcaaag
      901 tcagatgcag gatgccaggc agatccagcc atccccaccg tggtcctatg accagtccta
      961 ccagtacctg ggatccatca cctcttcctc tgtccaccca gcgacaccca tttcacccgg
     1021 ccgtgccagc ggcatgacca gcctctctgc agaactttcc agtcgactct caacggctcc
     1081 ggacctgacc gccttcggcg acccacgcca gttccctact ctgccgtcca tctccgaccc
     1141 gcgcatgcac tacccaggcg ccttcaccta ctcgccgccc gtcacgtcgg gcatcggcat
     1201 cggcatgtca gccatgagct cggcctctcg ctaccacacc tacctgccgc cgccctaccc
     1261 cggctcatca caggcgcagg ccgggccctt ccagaccggc tcgccctcct accatctata
     1321 ctacggcgcc tcggccggtt cctaccagtt ctccatggtg ggcggagaga gatcgccccc
     1381 gcgcatcctg ccgccctgca ccaacgcatc caccggcgcc gcgctgctca accccagcct
     1441 ccccagccag agcgacgtgg tggagaccga gggcagccat agcaactcgc ccaccaacat
     1501 gccccccgcg cgcctggagg aggccgtctg gcggccctac ctcgagtacc catacgacgt
     1561 cccagattac gcttgagaat tctgctatac gaagttatcc ctctccctcc ccccccccta
     1621 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     1681 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     1741 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     1801 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     1861 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1921 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1981 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     2041 taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca tgtgtttagt
     2101 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaagaa
     2161 cacgatgata atatggccac aaccatgact agtgccacca tggtgagcaa gggcgaggag
     2221 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     2281 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2341 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     2401 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     2461 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2521 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2581 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2641 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2701 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2761 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2821 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2881 accggcggca tggacgagct gtacaagtaa gcatcgataa aataaaagat tttatttagt
     2941 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     3001 acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc agatcaaggt
     3061 taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     3121 cccccggctc agggccaaga acagatggtc cccgatgcgg tcccgccctc agcagtttct
     3181 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     3241 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     3301 ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact gcgtcgcccg
     3361 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     3421 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catgggtaac
     3481 agtttcttga agttggagaa caacattctg agggtaggag tcgaatatta agtaatcctg
     3541 actcaattag ccactgtttt gaatccacat actccaatac tcctgaaata gttcattatg
     3601 gacagcgcag aagagctggg gagaattaat tcgtaatcat ggtcatagct gtttcctgtg
     3661 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3721 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3781 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3841 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3901 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3961 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4021 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4081 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4141 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4201 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4261 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4321 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4381 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4441 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4501 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4561 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4621 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4681 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4741 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4801 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4861 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4921 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4981 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5041 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5101 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5161 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5221 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5281 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5341 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5401 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5461 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5521 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5581 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5641 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5701 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5761 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5821 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     5881 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5941 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     6001 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     6061 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg
     6121 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     6181 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     6241 tgtaaaacga cggcgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     6301 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     6361 cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc
     6421 cggtgatgcc ggccacgatg cgtccggcgt agaggcgatt agtccaattt gttaaagaca
     6481 ggatatcagt ggtccaggct ctagttttga ctcaacaata tcaccagctg aagcctatag
     6541 agtacgagcc atagataaaa taaaagattt tatttagtct ccagaaaaag gggggaatga
     6601 aagaccccac ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga
     6661 aaatacataa ctgagaatag agaagttcag atcaaggtta ggaacagaga gacagcagaa
     6721 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     6781 gatggtcccc agatgcggtc ccgccctcag cagtttctag agaaccatca gatgtttcca
     6841 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     6901 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     6961 actcggcgcg ccagtcctcc gatagactgc gtcgcccggg tacccgtatt cccaataaag
     7021 cctcttgctg tttgcatccg aatcgtggac tcgctgatcc ttgggagggt ctcctcagat
     7081 tgattgactg cccacctcgg gggtctttca tttggaggtt ccaccgagat ttggagaccc
     7141 ctgcccaggg accaccgacc cccccgccgg gaggtaagct ggccagcggt cgtttcgtgt
     7201 ctgtctctgt ctttgtgcgt gtttgtgccg gcatctaatg tttgcgcctg cgtctgtact
     7261 agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag ttctgaacac
     7321 ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt ggcccgacct
     7381 gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg gtaggagacg
     7441 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg aaccgaagcc
     7501 gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt
     7561 ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag
     7621 gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac
     7681 gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg
     7741 gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc
     7801 atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc
     7861 ctccctgggt ca
//
