LOCUS       Exported                7089 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  retroviral expression of Runx1DN and mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    Banshee RunxDN mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7089)
  TITLE     Rothenberg lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7089)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7089
                     /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             764..790
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    821..1388
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    838..1390
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(982..999)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1209..1228
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     regulatory      1411..1420
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1417..2127
                     /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(1562..1580)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1830..1849
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     LTR             2176..2690
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     primer_bind     complement(2859..2875)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2859..2875)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2872..2894)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2883..2899
                     /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(2907..2937)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2952..2973
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3090..3107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3261..3849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3341..3360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4020..4880)
                     /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     4643..4662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4881..4985)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5053..5071
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5109..5131)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5231..5250
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5444..5466
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5594..5613
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             5813..6329
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    6393..6734
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             6801..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 gcccgggctc gagtacccat acgacgtccc
      781 agattacgct tgagaattct gctatacgaa gttatccctc tccctccccc ccccctaacg
      841 ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca
      901 ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga
      961 gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga
     1021 aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca
     1081 ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag
     1141 atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa
     1201 gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac
     1261 cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt gtttagtcga
     1321 ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaagaacac
     1381 gatgataata tggccacaac catgactagt gccaccatgg tgagcaaggg cgaggaggat
     1441 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
     1501 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
     1561 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
     1621 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
     1681 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
     1741 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
     1801 ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg
     1861 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     1921 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     1981 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     2041 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     2101 ggcggcatgg acgagctgta caagtaagca tcgataaaat aaaagatttt atttagtctc
     2161 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     2221 ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga tcaaggttag
     2281 gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     2341 ccggctcagg gccaagaaca gatggtcccc gatgcggtcc cgccctcagc agtttctaga
     2401 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     2461 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     2521 aaagagccca caacccctca ctcggcgcgc cagtcctccg atagactgcg tcgcccgggt
     2581 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     2641 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcat gggtaacagt
     2701 ttcttgaagt tggagaacaa cattctgagg gtaggagtcg aatattaagt aatcctgact
     2761 caattagcca ctgttttgaa tccacatact ccaatactcc tgaaatagtt cattatggac
     2821 agcgcagaag agctggggag aattaattcg taatcatggt catagctgtt tcctgtgtga
     2881 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2941 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3001 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3061 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3121 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3181 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3241 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3301 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3361 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3421 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3481 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3541 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3601 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3661 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3721 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3781 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3841 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3901 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3961 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4021 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4081 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4141 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4201 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4261 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4321 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4381 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4441 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4501 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4561 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4621 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4681 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4741 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4801 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4861 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4921 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4981 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     5041 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     5101 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     5161 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     5221 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     5281 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     5341 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     5401 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     5461 aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     5521 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     5581 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     5641 tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt ccaatttgtt aaagacagga
     5701 tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag cctatagagt
     5761 acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg ggaatgaaag
     5821 accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa
     5881 tacataactg agaatagaga agttcagatc aaggttagga acagagagac agcagaatat
     5941 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     6001 ggtccccaga tgcggtcccg ccctcagcag tttctagaga accatcagat gtttccaggg
     6061 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     6121 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     6181 cggcgcgcca gtcctccgat agactgcgtc gcccgggtac ccgtattccc aataaagcct
     6241 cttgctgttt gcatccgaat cgtggactcg ctgatccttg ggagggtctc ctcagattga
     6301 ttgactgccc acctcggggg tctttcattt ggaggttcca ccgagatttg gagacccctg
     6361 cccagggacc accgaccccc ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg
     6421 tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt
     6481 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg
     6541 gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag
     6601 gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga
     6661 acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg
     6721 cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc
     6781 tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc
     6841 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     6901 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     6961 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     7021 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     7081 cctgggtca
//
