LOCUS       Exported                8204 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 8204)
  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..8204
                     /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 1-465aa (soluble) Y213A/I216A W276A/I279A
                     /translation="MASSSTVPLGFHYETKYVVLSYLGLLSQEKLQEQHLSSPQGVQLD
                     IASQSLDQEILLKVKTEIEEELKSLDKEISEAFTSTGFDRHTSPVFSPANPESSMEDCL
                     AHLGEKVSQELKEPLHKALQMLLSQPVTYQAFRECTLETTVHASGWNKILVPLVLLRQM
                     LLELTRRGQEPLSALLQFGVTYLEDYSAEYIIQQGGWGTVFSLESEEEEAPGATAEDSN
                     DIYILPSDNSGQVSPPESPTVTTSWQSESLPVSLSASQSWHTESLPVSLGPESAQQAAM
                     DPEEVKSLDSNGAGEKSENNSSNSDIVHVEKEEVPEGMEEAAVASVVLPARELQEALPE
                     APAPLLPHITATSLLGTREPDTEVITVEKSSPATSLFVELDEEEVKAATTEPTEVEEVV
                     PALEPTETLLSEKEINAREESLVEELSPASEKKPVPPSEGKSRLSPAGEMKPMPLSEGK
                     SILLFGG"
     primer_bind     300..320
                     /label=Fragment.FOR (1)
     misc_feature    936..947
                     /label=FIR2mut
     misc_feature    969..980
                     /label=FIR3
     misc_feature    1125..1136
                     /label=LIR1mut
     misc_feature    1218..1229
                     /label=FIR4
     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     5911..5930
                     /label=GEX3-rev
     primer_bind     5940..5958
                     /label=GEX3-rev-25
     primer_bind     5972..5991
                     /label=SMP3644
     primer_bind     complement(6717..6734)
                     /label=SMP2212
     CDS             complement(6718..7674)
                     /label=LacI
     primer_bind     7878..7896
                     /label=SMP52
     primer_bind     8043..8060
                     /label=SMP2285
     primer_bind     8126..8145
                     /label=SMP2293
     primer_bind     8142..8157
                     /label=SMP49
     primer_bind     8142..8156
                     /label=pET-up
     primer_bind     8170..8186
                     /label=SMP1677
     primer_bind     8177..8194
                     /label=T7probis
     primer_bind     join(8188..8204,1..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 aggaagctcc tggagctact gcagaagata
      961 gcaatgacat ttacatcctg cccagcgaca actctggaca agtcagtccc ccagagtctc
     1021 caactgtgac cacttcctgg cagtctgaga gcttacctgt gtcactgtca gctagccaga
     1081 gttggcacac agaaagcctg ccagtgtcac taggccctga gtccgcgcag caggctgcaa
     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 caccgcatat
     5761 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     5821 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     5881 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     5941 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     6001 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     6061 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     6121 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     6181 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     6241 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     6301 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     6361 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     6421 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     6481 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     6541 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     6601 ctagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     6661 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     6721 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6781 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     6841 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     6901 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     6961 gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga
     7021 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     7081 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     7141 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     7201 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     7261 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     7321 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     7381 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     7441 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     7501 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     7561 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     7621 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     7681 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     7741 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     7801 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     7861 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     7921 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     7981 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     8041 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     8101 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     8161 tcgagatcga tctcgatccc gcgaaattaa tacgactcac tata
//
