LOCUS       Exported                9029 bp DNA     circular SYN 13-APR-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pLego.sgGata1.4.RUNX1A.iG2_approved
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9029)
  AUTHORS   Universitaetsklinikum Halle
  TITLE     Direct Submission
  JOURNAL   Exported Apr 26, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9029
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
                     /note="color: #ffe4c4"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
                     /note="color: #ffe4c4"
     misc_feature    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
                     /note="color: #ffe4c4"
     promoter        2620..2933
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2935..2955
                     /label=sg Gata1.4
                     /note="color: #ff0000"
     protein_bind    3107..3140
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     promoter        3211..3617
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
                     /note="color: #ffffff; direction: RIGHT"
     regulatory      3622..3631
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             3634..3660
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="color: #cc99b2"
                     /translation="YPYDVPDYA"
     CDS             3661..4410
                     /codon_start=1
                     /label=RUNX1A
                     /note="color: #ff0000"
                     /translation="RIPVDASTSRRFTPPSTALSPGKMSEALPLGAPDAGAALAGKLRS
                     GDRSMVEVLADHPGELVRTDSPNFLCSVLPTHWRCNKTLPIAFKVVALGDVPDGTLVTV
                     MAGNDENYSAELRNATAAMKNQVARFNDLRFVGRSGRGKSFTLTITVFTNPPQVATYHR
                     AIKITVDGPREPRRHRQKLDDQTKPGSLSFSERLSELEQLRRTAMRVSPHHPAPTPNPR
                     ASLNHSTAFNPQPQSQMQEEDTAPWRC"
     misc_feature    4482..5049
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="color: #a6acb3"
     CDS             5063..5782
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 5063 .. 5065 / #05fd14
                        2: 5066 .. 5068 / #05fd14 / 1a
                        3: 5069 .. 5782 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    5829..5862
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    5918..6506
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
                     /note="color: #ffffff"
     CDS             complement(6389..6400)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /note="color: #cc99b2
                     Cleavage site after base 6388"
                     /translation="IEGR"
     LTR             7035..7215
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /note="color: #ffe4c4"
     rep_origin      complement(7277..7865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(8036..8896)
                     /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: 8036 .. 8827 / #ccffcc
                        2: 8828 .. 8896 / #ccffcc / signal sequence
                     Cleavage site after base 8827"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8897..9001)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgctctagag atccgacgcc gccatctcta
     2641 ggcccgcgcc ggccccctcg cacagacttg tgggagaagc tcggctactc ccctgccccg
     2701 gttaatttgc atataatatt tcctagtaac tatagaggct taatgtgcga taaaagacag
     2761 ataatctgtt ctttttaata ctagctacat tttacatgat aggcttggat ttctataaga
     2821 gatacaaata ctaaattatt attttaaaaa acagcacaaa aggaaactca ccctaactgt
     2881 aaagtaattg tgtgttttga gactataaat atcccttgga gaaaagcctt gtttgcctag
     2941 accaggaaaa tccatgttta agagctatgc tggaaacagc atagcaagtt taaataaggc
     3001 tagtccgtta tcaacttgaa aaagtggcac cgagtcggtg ctttttctcg aggtcgacgg
     3061 tatcgataag ctcgcttcac gagattccag caggtcgagg gacctaataa cttcgtatag
     3121 catacattat acgaagttat attaagggtt ccaagcttaa gcggccggcc gctgaaagac
     3181 cccacctgta ggtttggcaa gctagctgca gtaacgccat tttgcaaggc atggaaaaat
     3241 accaaaccaa gaatagagaa gttcagatca agggcgggta catgaaaata gctaacgttg
     3301 ggccaaacag gatatctgcg gtgagcagtt tcggccccgg cccggggcca agaacagatg
     3361 gtcaccgcag tttcggcccc ggcccgaggc caagaacaga tggtccccag atatggccca
     3421 accctcagca gtttcttaag acccatcaga tgtttccagg ctcccccaag gacctgaaat
     3481 gaccctgcgc cttatttgaa ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt
     3541 ctgcttcccg agctctataa aagagctcac aacccctcac tcggcgcgcc agtcctccga
     3601 ttgactgagt cgcccggatc cgccaccatg ggatacccct acgacgtgcc cgactacgcc
     3661 cgtatccccg tagatgccag cacgagccgc cgcttcacgc cgccttccac cgcgctgagc
     3721 ccaggcaaga tgagcgaggc gttgccgctg ggcgccccgg acgccggcgc tgccctggcc
     3781 ggcaagctga ggagcggcga ccgcagcatg gtggaggtgc tggccgacca cccgggcgag
     3841 ctggtgcgca ccgacagccc caacttcctc tgctccgtgc tgcctacgca ctggcgctgc
     3901 aacaagaccc tgcccatcgc tttcaaggtg gtggccctag gggatgttcc agatggcact
     3961 ctggtcactg tgatggctgg caatgatgaa aactactcgg ctgagctgag aaatgctacc
     4021 gcagccatga agaaccaggt tgcaagattt aatgacctca ggtttgtcgg tcgaagtgga
     4081 agagggaaaa gcttcactct gaccatcact gtcttcacaa acccaccgca agtcgccacc
     4141 taccacagag ccatcaaaat cacagtggat gggccccgag aacctcgaag acatcggcag
     4201 aaactagatg atcagaccaa gcccgggagc ttgtcctttt ccgagcggct cagtgaactg
     4261 gagcagctgc ggcgcacagc catgagggtc agcccacacc acccagcccc cacgcccaac
     4321 cctcgtgcct ccctgaacca ctccactgcc tttaaccctc agcctcagag tcagatgcag
     4381 gaggaagaca cagcaccctg gagatgttaa gaattcctgc aggcctcgac gagggccggc
     4441 gcgccgcggc cgctacgtaa attccgcccc ccccccccny ctccctcccc cccccctaac
     4501 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     4561 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     4621 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     4681 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     4741 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     4801 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     4861 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     4921 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     4981 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     5041 cgatgataat atggccacaa ccatggtgag caagggcgag gagctgttca ccggggtggt
     5101 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     5161 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     5221 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     5281 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     5341 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5401 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5461 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     5521 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     5581 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     5641 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     5701 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     5761 catggacgag ctgtacaagt aaagcggccg gccgccagca cagtggtcga aattcgtcga
     5821 gggacctaat aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg
     5881 gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     5941 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     6001 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     6061 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     6121 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     6181 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     6241 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     6301 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     6361 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     6421 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     6481 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
     6541 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     6601 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     6661 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     6721 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     6781 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
     6841 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
     6901 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     6961 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
     7021 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     7081 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     7141 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     7201 ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7261 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7321 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7381 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7441 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     7501 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7561 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7621 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7681 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     7741 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7801 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7861 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7921 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7981 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8041 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8101 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8161 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8221 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8281 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8341 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     8401 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8461 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8521 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8581 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8641 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8701 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8761 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8821 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8881 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8941 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9001 gcacatttcc ccgaaaagtg ccacctgac
//
