LOCUS       Exported                7181 bp ds-DNA     circular SYN 16-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7181)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 30, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7181
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(259..363)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             393..986
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    1049..1406
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1465..1881
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     misc_feature    1891..2265
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     regulatory      2294..2303
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2303..2368
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2402..3115
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     misc_feature    3140..3862
                     /label=PLEKHB1_Homo_sapiens
     misc_feature    3907..4490
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     LTR             5057..5526
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(5827..6415)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
       61 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      361 gcgcacattt ccccgaaaag tgccacctgc ataatgaaag accccacctg taggtttggc
      421 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagaa
      481 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
      541 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
      601 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
      661 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccggggtgc
      721 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
      781 ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg
      841 cgcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt
      901 gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga
      961 ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc
     1021 agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg
     1081 attgtctagt gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc
     1141 tctgtatctg gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg
     1201 ggagacgtcc cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc
     1261 gatcgttttg gactctttgg tgcacccccc taataggagg gatatgtggt tctggtagga
     1321 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga
     1381 agccgcgccg cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg
     1441 tttctgtatt tgtctgaaaa ttagggccag actgttacca ctcccttaag tttgacctta
     1501 ggtcactgga aagatgtcga gcggatcgct cacaaccagt cggtagatgt caagaagaga
     1561 cgttgggtta ccttctgctc tgcagaatgg ccaaccttta acgtcggatg gccgcgagac
     1621 ggcaccttta accgagacct catcacccag gttaagatca aggtcttttc acctggcccg
     1681 catggacacc cagaccaggt cccctacatc gtgacctggg aagccttggc ttttgacccc
     1741 cctccctggg tcaagccctt tgtacaccct aagcctccgc ctcctcttcc tccatccgcc
     1801 ccgtctctcc cccttgaacc tcctcgttcg accccgcctc gatcctccct ttatccagcc
     1861 ctcactcctt ctctaggcgc ccccatatgg ccatatgaga tcttatatgg ggcacccccg
     1921 ccccttgtaa acttccctga ccctgacatg acaagagtta ctaacagccc ctctctccaa
     1981 gctcacttac aggctctcta cttagtccag cacgaagtct ggagacctct ggcggcagcc
     2041 taccaagaac aactggaccg accggtggta cctcaccctt accgagtcgg cgacacagtg
     2101 tgggtccgcc gacaccagac taagaaccta gaacctcgct ggaaaggacc ttacacagtc
     2161 ctgctgacca cccccaccgc cctcaaagta gacggcatcg cagcttggat acacgccgcc
     2221 cacgtgaagg ctgccgaccc cgggggtgga ccatcctcta gactgccgga tctagctagt
     2281 taattaagga tccgccacca tggactacaa ggaccacgac ggtgactaca aggaccacga
     2341 catcgactac aaggacgacg acgacaaggg gcatggcacc ggcagcaccg gcagcgaatt
     2401 cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2461 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     2521 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2581 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2641 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2701 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2761 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2821 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2881 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2941 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     3001 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     3061 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg
     3121 gagcggcccg cggctcgaga tgtgggtgga gggtcctcat tcagatgccc caggcctcgg
     3181 atcctgggcg ggggcgcgtg gcagccgcca ggagaccggc tggagcgccc gccccgcggc
     3241 ctcgcctctc ctccgagcag ccagcgcctc gggacgcgat gaggaccttg gcttgcctgc
     3301 tgctcctcgg ctgcggatac ctcgcccatg ttctggccga ggaagccgag atcccccgcg
     3361 aggtgatcga gaggctggcc cgcagtcaga tccacagcat ccgggacctc cagcgactcc
     3421 tggagataga ctccggagtg aggattcttt ggacaccagc ctgagagctc acggggtcca
     3481 tgccactaag catgtgcccg agaagcggcc cctgcccatt cggaggaaga gaagcatcga
     3541 ggaagctgtc cccgctgtct gcaagaccag gacggtcatt tacgagattc ctcggagtca
     3601 ggtcgacccc acgtccgcca acttcctgat ctggcccccg tgcgtggagg tgaaacgctg
     3661 caccggctgc tgcaacacga gcagtgtcaa gtgccagccc tcccgcgtcc accaccgcag
     3721 cgtcaaggtg gccaaggtgg aatacgtcag gaagaagcca aaattaaaag aagtccaggt
     3781 gaggttagag gagcatttgg agtgcgcctg cgcgaccaca agcctgaatc cggattatcg
     3841 ggaagaggac acggatgtga ggtgagcggc cgctaagcag gccgctacgt aaattccgcc
     3901 cccccccccc tctccctccc cccctaacgt tactggccga agccgcttgg aataaggccg
     3961 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     4021 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     4081 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     4141 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     4201 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     4261 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     4321 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     4381 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     4441 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc atggttcctt
     4501 tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca
     4561 tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg
     4621 gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg
     4681 ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg
     4741 gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca
     4801 aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc
     4861 cctctggtta tgtgtgggag ggctaagtcg acgataaaat aaaagatttt atttagtctc
     4921 cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc ttaagtaacg
     4981 ccattttgca aggcatggaa aaatacataa ctgagaatag agaagttcag atcaaggtca
     5041 ggaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     5101 cccggctcag ggccaagaac agatggaaca gctgaatatg ggccaaacag gatatctgtg
     5161 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtccagc
     5221 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     5281 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     5341 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggggcgccag tcctccgatt
     5401 gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     5461 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     5521 ctttcacatg cagcatgtat caaaattaat ttggtttttt ttcttaagta tttacattaa
     5581 atggccatag ttgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5641 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5701 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5761 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5821 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5881 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5941 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6001 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6061 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6121 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6181 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6241 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6301 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6361 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6421 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6481 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttgcggc cggccgcaaa
     6541 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6601 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6661 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6721 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6781 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6841 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6901 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6961 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7021 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7081 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7141 aagtcattct gagaatagtg tatgcggcga ccgagttgct c
//
