LOCUS       Exported                8201 bp ds-DNA     circular SYN 01-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8201)
  AUTHORS   eliasa55
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8201
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_binding    3..25
                     /label=LacO
     primer_bind     45..61
                     /label=SMP2196
     CDS             83..100
                     /label=6His
     primer_bind     83..100
                     /label=SMP3647
     primer_bind     complement(97..124)
                     /label=SMP3044
     primer_bind     189..208
                     /label=SMP50
     primer_bind     complement(189..208)
                     /label=SMP51
     primer_bind     complement(191..208)
                     /label=SMP3659
     primer_bind     214..233
                     /label=T7
     misc_binding    233..255
                     /label=LacO
     primer_bind     complement(254..299)
                     /label=Vector.REV
     primer_bind     complement(261..315)
                     /label=Vector.REV (2)
     primer_bind     275..321
                     /label=Fragment.FOR
     CDS             300..1694
                     /codon_start=1
                     /label=BCL2L13 1-465 (soluble)
                     /translation="MASSSTVPLGFHYETKYVVLSYLGLLSQEKLQEQHLSSPQGVQLD
                     IASQSLDQEILLKVKTEIEEELKSLDKEISEAFTSTGFDRHTSPVFSPANPESSMEDCL
                     AHLGEKVSQELKEPLHKALQMLLSQPVTYQAFRECTLETTVHASGWNKILVPLVLLRQM
                     LLELTRRGQEPLSALLQFGVTYLEDYSAEYIIQQGGWGTVFSLESEEEEYPGITAEDSN
                     DIYILPSDNSGQVSPPESPTVTTSWQSESLPVSLSASQSWHTESLPVSLGPESWQQIAM
                     DPEEVKSLDSNGAGEKSENNSSNSDIVHVEKEEVPEGMEEAAVASVVLPARELQEALPE
                     APAPLLPHITATSLLGTREPDTEVITVEKSSPATSLFVELDEEEVKAATTEPTEVEEVV
                     PALEPTETLLSEKEINAREESLVEELSPASEKKPVPPSEGKSRLSPAGEMKPMPLSEGK
                     SILLFGG"
     primer_bind     300..320
                     /label=Fragment.FOR (1)
     primer_bind     complement(1669..1714)
                     /label=Fragment.REV (1)
     primer_bind     complement(1669..1694)
                     /label=Fragment.REV
     primer_bind     1687..1724
                     /label=Vector.FOR (2)
     CDS             1707..2420
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=mEGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     1707..1724
                     /label=Fragment.FORa
     primer_bind     complement(2432..2462)
                     /label=Fragment.REVa
     CDS             2433..2453
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     2440..2502
                     /label=SMP2156
     primer_bind     2447..2473
                     /label=Vector.FOR (1)
     primer_bind     2454..2472
                     /label=Vector.FOR
     primer_bind     complement(2454..2468)
                     /label=Fragment.REV
     CDS             2460..2477
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    2484..3134
                     /label=GST no Met
     primer_bind     complement(2524..2541)
                     /label=SMP2192
     primer_bind     complement(2780..2803)
                     /label=SMP1669
     primer_bind     3019..3040
                     /label=SMP358
     primer_bind     3064..3083
                     /label=GEX5-for
     primer_bind     3095..3112
                     /label=pETG-for
     primer_bind     3096..3115
                     /label=SMP3299
     CDS             3150..3194
                     /label=S-tag
     primer_bind     complement(3150..3166)
                     /label=SMP2284
     primer_bind     complement(3232..3251)
                     /label=T7term
     primer_bind     complement(3232..3250)
                     /label=SMP545
     primer_bind     complement(3273..3288)
                     /label=T7terbis
     primer_bind     complement(3293..3313)
                     /label=pGMP-rev
     rep_origin      3330..3785
                     /direction=RIGHT
                     /label=M13 origin
     rep_origin      complement(3335..3775)
                     /direction=LEFT
                     /label=F1 ori
     CDS             complement(3903..4562)
                     /label=AmpR
     primer_bind     complement(3950..3969)
                     /label=AmpStop
     primer_bind     4696..4715
                     /label=AmpStart
     promoter        complement(4803..4831)
                     /label=Amp prom
     rep_origin      4912..5540
                     /direction=RIGHT
                     /label=ColE1 origin
     primer_bind     5908..5927
                     /label=GEX3-rev
     primer_bind     5937..5955
                     /label=GEX3-rev-25
     primer_bind     5969..5988
                     /label=SMP3644
     primer_bind     complement(6714..6731)
                     /label=SMP2212
     CDS             complement(6715..7671)
                     /label=LacI
     primer_bind     7875..7893
                     /label=SMP52
     primer_bind     8040..8057
                     /label=SMP2285
     primer_bind     8123..8142
                     /label=SMP2293
     primer_bind     8139..8154
                     /label=SMP49
     primer_bind     8139..8153
                     /label=pET-up
     primer_bind     8167..8183
                     /label=SMP1677
     primer_bind     8174..8191
                     /label=T7probis
     primer_bind     join(8185..8201,1..3)
                     /label=T7
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt
      241 gagcggataa caattcccca tcttagtata ttagttaagt ataagaagga gatatacata
      301 tggcgtcctc ttctactgtg cctctgggat ttcactatga aacaaagtat gttgttctca
      361 gctacttggg actcctctct caagagaagc tgcaagagca acatctttcc tcaccccaag
      421 gggttcaact agatatagct tcacaatctc tggatcaaga aattttatta aaagttaaaa
      481 ctgaaattga agaagagcta aaatctctgg acaaagaaat ttctgaagcc ttcaccagca
      541 caggctttga ccgtcacact tctccagtgt tcagccctgc caatccagaa agctcaatgg
      601 aagactgctt ggcccatctt ggagaaaaag tgtcccagga actgaaagag cctctccata
      661 aagcattgca aatgctcctg agccagccag tgacatatca ggcatttcgg gaatgtacac
      721 tggagaccac agttcatgcc agcggctgga ataagatttt ggtgcctctg gttttgctac
      781 gacaaatgct tttggaattg acaagacgtg gtcaagaacc tttgagcgca ctgctgcagt
      841 ttggcgtgac atacctggag gactattcgg cagagtacat cattcagcaa ggtggctggg
      901 gcactgtgtt tagtcttgag tcagaggagg aggaataccc tggaatcact gcagaagata
      961 gcaatgacat ttacatcctg cccagcgaca actctggaca agtcagtccc ccagagtctc
     1021 caactgtgac cacttcctgg cagtctgaga gcttacctgt gtcactgtca gctagccaga
     1081 gttggcacac agaaagcctg ccagtgtcac taggccctga gtcctggcag cagattgcaa
     1141 tggatcctga agaagtgaaa agcttagaca gcaacggagc tggagagaag agtgagaaca
     1201 actcctctaa ttctgacatt gtgcacgtgg agaaagaaga ggtgcccgag ggcatggaag
     1261 aggctgctgt ggcttctgtg gtcttgccag cgcgggagct gcaagaggca cttcctgaag
     1321 ccccagctcc cttgcttcca catatcactg ccacctccct gctggggaca agggaacctg
     1381 acacagaagt gatcacagtt gagaaatcca gccctgctac atctctgttt gtagaacttg
     1441 atgaagaaga ggtgaaagca gcaacaactg aacctactga agtggaggag gtggtccccg
     1501 cactggaacc cacagaaacg ctgctgagtg agaaggagat aaacgcaagg gaagagagcc
     1561 ttgtggaaga gctgtcccct gccagcgaga agaagcccgt gccgccgtct gagggcaagt
     1621 ctagactgtc ccccgccggt gagatgaagc ccatgccgct gtctgagggc aagtctatac
     1681 tgctgtttgg aggaggcggc ggcggcgtga gcaagggcga ggagctgttc accggggtgg
     1741 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1801 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1861 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1921 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1981 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     2041 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     2101 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     2161 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     2221 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     2281 ccgtgctgct gcccgacaac cactacctga gcacccagtc caagctgagc aaagacccca
     2341 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     2401 gcatggacga gctgtacaag ggcggcagtg gcgaaaacct gtattttcag ggcggatcac
     2461 tggttccgcg tggatccggc ggatccccta tactaggtta ttggaaaatt aagggccttg
     2521 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
     2581 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
     2641 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
     2701 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
     2761 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
     2821 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
     2881 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca
     2941 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
     3001 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
     3061 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
     3121 accatcctcc aaaatagctc gagtctggta aagaaaccgc tgctgcgaaa tttgaacgcc
     3181 agcacatgga ctcgtctact agcgcagctt aattaaccta ggctgctgcc accgctgagc
     3241 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     3301 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     3361 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3421 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3481 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3541 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3601 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3661 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3721 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
     3781 aatttctggc ggcacgatgg catgagatta tcaaaaagga tcttcaccta gatcctttta
     3841 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3901 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3961 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4021 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4081 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4141 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4201 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4261 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4321 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4381 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4441 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4501 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4561 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4621 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4681 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4741 cggaaatgtt gaatactcat actcttcctt tttcaatcat gattgaagca tttatcaggg
     4801 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggtca
     4861 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     4921 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     4981 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5041 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5101 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5161 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5221 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5281 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5341 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5401 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5461 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     5521 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     5581 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     5641 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     5701 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg
     5761 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta
     5821 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     5881 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5941 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc
     6001 tcatcagcgt ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg
     6061 ttgagtttct ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg
     6121 gttttttcct gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta
     6181 atgataccga tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc
     6241 cggttactgg aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga
     6301 aaaatcactc agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt
     6361 agccagcagc atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc
     6421 gtttccagac tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca
     6481 gacgttttgc agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa
     6541 ccagtaaggc aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcta
     6601 gtcatgcccc gcgcccaccg gaaggagctg actgggttga aggctctcaa gggcatcggt
     6661 cgagatcccg gtgcctaatg agtgagctaa cttacattaa ttgcgttgcg ctcactgccc
     6721 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6781 agaggcggtt tgcgtattgg gcgccagggt ggtttttctt ttcaccagtg agacgggcaa
     6841 cagctgattg cccttcaccg cctggccctg agagagttgc agcaagcggt ccacgctggt
     6901 ttgccccagc aggcgaaaat cctgtttgat ggtggttaac ggcgggatat aacatgagct
     6961 gtcttcggta tcgtcgtatc ccactaccga gatgtccgca ccaacgcgca gcccggactc
     7021 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
     7081 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
     7141 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
     7201 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
     7261 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
     7321 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
     7381 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
     7441 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
     7501 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
     7561 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
     7621 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
     7681 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
     7741 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
     7801 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
     7861 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
     7921 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     7981 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     8041 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     8101 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     8161 agatcgatct cgatcccgcg aaattaatac gactcactat a
//
