LOCUS       Exported                7898 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 7898)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Sep 30, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7898
                     /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..4579
                     /label=EDIL3_Homo_sapiens
     misc_feature    4624..5207
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     LTR             5774..6243
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     rep_origin      complement(6544..7132)
                     /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 tgaagcgctc ggtagccgtc tggctcttgg tcgggctcag
     3181 cctcggtgtc ccccagttcg gcaaaggtga tatttgtgat cccaatccat gtgaaaatgg
     3241 aggtatctgt ttgccaggat tggctgatgg ttccttttcc tgtgagtgtc cagatggctt
     3301 cacagacccc aactgttcta gtgttgtgga ggttgcatca gatgaagaag aaccaacttc
     3361 agcaggtccc tgcactccta atccatgcca taatggagga acctgtgaaa taagtgaagc
     3421 ataccgaggg gatacattca taggctatgt ttgtaaatgt ccccgaggat ttaatgggat
     3481 tcactgtcag cacaacataa atgaatgcga agttgagcct tgcaaaaatg gtggaatatg
     3541 tacagatctt gttgctaact attcctgtga gtgcccaggc gaatttatgg gaagaaattg
     3601 tcaatacaaa tgctcaggcc cactgggaat tgaaggtgga attatatcaa accagcaaat
     3661 cacagcttcc tctactcacc gagctctttt tggactccaa aaatggtatc cctactatgc
     3721 acgtcttaat aagaaggggc ttataaatgc gtggacagct gcagaaaatg acagatggcc
     3781 gtggattcag ataaatttgc aaaggaaaat gagagttact ggtgtgatta cccaaggagc
     3841 caagaggatt ggaagcccag agtatataaa atcctacaaa attgcctaca gtaatgatgg
     3901 aaagacttgg gcaatgtaca aagtgaaagg caccaatgaa gacatggtgt ttcgtggaaa
     3961 cattgataac aacactccat atgctaactc tttcacaccc cccataaaag ctcagtatgt
     4021 aagactctat ccccaagttt gtcgaagaca ttgcactttg cgaatggaac ttcttggctg
     4081 tgaactgtcg ggttgttctg agcctctggg tatgaaatca ggacatatac aagactatca
     4141 gatcactgcc tccagcatct tcagaacgct caacatggac atgttcactt gggaaccaag
     4201 gaaagctcgg ctggacaagc aaggcaaagt gaatgcctgg acctctggcc acaatgacca
     4261 gtcacaatgg ttacaggtgg atcttcttgt tccaaccaaa gtgactggca tcattacaca
     4321 aggagctaaa gattttggtc atgtacagtt tgttggctcc tacaaactgg cttacagcaa
     4381 tgatggagaa cactggactg tataccagga tgaaaagcaa agaaaagata aggttttcca
     4441 gggaaatttt gacaatgaca ctcacagaaa aaatgtcatc gaccctccca tctatgcacg
     4501 acacataaga atccttcctt ggtcctggta cgggaggatc acattgcggt cagagctgct
     4561 gggctgcaca gaggaggaat gagcggccgc taagcaggcc gctacgtaaa ttccgccccc
     4621 ccccccctct ccctcccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg
     4681 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg
     4741 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg
     4801 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca
     4861 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct
     4921 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca
     4981 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa
     5041 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg
     5101 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg
     5161 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gttcctttgt
     5221 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct
     5281 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg
     5341 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg
     5401 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca
     5461 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag
     5521 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct
     5581 ctggttatgt gtgggagggc taagtcgacg ataaaataaa agattttatt tagtctccag
     5641 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     5701 ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga
     5761 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     5821 ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     5881 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct
     5941 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     6001 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     6061 ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
     6121 tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     6181 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     6241 tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg
     6301 gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6361 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6421 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6481 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6541 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6601 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6661 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6721 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6781 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6841 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6901 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6961 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     7021 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     7081 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     7141 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     7201 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgg ccgcaaatca
     7261 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     7321 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     7381 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     7441 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     7501 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7561 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7621 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7681 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7741 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7801 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7861 tcattctgag aatagtgtat gcggcgaccg agttgctc
//
