LOCUS       Exported                6953 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 6953)
  AUTHORS   Max Perutz Labs
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 8, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6953
                     /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(258..309)
                     /label=SMP4627
                     /note="reverse primer to linearize SMC2045 for insertion of
                     FUNDC1 sequence (1-50aa)"
     primer_bind     291..315
                     /label=SMP4624
                     /note="forward primer to insert FUNDC1 sequence into 
                     SMC2045 (instead of BCL2L13 insert)"
     CDS             300..449
                     /codon_start=1
                     /label=soluble FUNDC1 (1-50)
                     /translation="MATRNPPPQDYESDDDSYEVLDLTEYARRHQWWNRVFGHSSGPMV
                     EKYSV"
     misc_feature    351..362
                     /label=LIR
     primer_bind     complement(418..457)
                     /label=SMP4625
                     /note="reverse primer to insert FUNDC1 sequence into 
                     SMC2045 (instead of BCL2L13 insert)"
     primer_bind     complement(420..451)
                     /label=Fragment.REV
     primer_bind     443..464
                     /label=SMP4626
                     /note="forward primer to linearize SMC2045 for insertion of
                     FUNDC1 sequence (1-50aa)"
     CDS             456..1169
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=mEGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     456..473
                     /label=Fragment.FORa
     misc_feature    1071..1073
                     /label=A206K
                     /note="monomeric EGFP"
     CDS             1182..1202
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     1189..1251
                     /label=SMP2156
     primer_bind     1203..1221
                     /label=Vector.FOR
     primer_bind     complement(1203..1217)
                     /label=Fragment.REV
     CDS             1209..1226
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     misc_feature    1233..1883
                     /label=GST no Met
     primer_bind     complement(1273..1290)
                     /label=SMP2192
     primer_bind     complement(1529..1552)
                     /label=SMP1669
     primer_bind     1768..1789
                     /label=SMP358
     primer_bind     1813..1832
                     /label=GEX5-for
     primer_bind     1844..1861
                     /label=pETG-for
     primer_bind     1845..1864
                     /label=SMP3299
     CDS             1899..1943
                     /label=S-tag
     primer_bind     complement(1899..1915)
                     /label=SMP2284
     primer_bind     complement(1981..2000)
                     /label=T7term
     primer_bind     complement(1981..1999)
                     /label=SMP545
     primer_bind     complement(2022..2037)
                     /label=T7terbis
     primer_bind     complement(2042..2062)
                     /label=pGMP-rev
     rep_origin      2079..2534
                     /direction=RIGHT
                     /label=M13 origin
     rep_origin      complement(2084..2524)
                     /direction=LEFT
                     /label=F1 ori
     CDS             complement(2652..3311)
                     /label=AmpR
     primer_bind     complement(2699..2718)
                     /label=AmpStop
     primer_bind     3445..3464
                     /label=AmpStart
     promoter        complement(3552..3580)
                     /label=Amp prom
     rep_origin      3661..4289
                     /direction=RIGHT
                     /label=ColE1 origin
     primer_bind     4660..4679
                     /label=GEX3-rev
     primer_bind     4689..4707
                     /label=GEX3-rev-25
     primer_bind     4721..4740
                     /label=SMP3644
     primer_bind     complement(5466..5483)
                     /label=SMP2212
     CDS             complement(5467..6423)
                     /label=LacI
     primer_bind     6627..6645
                     /label=SMP52
     primer_bind     6792..6809
                     /label=SMP2285
     primer_bind     6875..6894
                     /label=SMP2293
     primer_bind     6891..6906
                     /label=SMP49
     primer_bind     6891..6905
                     /label=pET-up
     primer_bind     6919..6935
                     /label=SMP1677
     primer_bind     6926..6943
                     /label=T7probis
     primer_bind     join(6937..6953,1..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 tggcgacccg gaacccccct ccccaagact atgaaagtga tgacgactct tatgaagtgt
      361 tggatttaac tgagtatgca agaagacacc agtggtggaa tcgagtgttt ggccacagtt
      421 cgggacctat ggtagaaaaa tactcagtag gcggcgtgag caagggcgag gagctgttca
      481 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      541 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      601 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      661 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      721 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      781 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      841 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      901 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
      961 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1021 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca
     1081 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1141 tcactctcgg catggacgag ctgtacaagg gcggcagtgg cgaaaacctg tattttcagg
     1201 gcggatcact ggttccgcgt ggatccggcg gatcccctat actaggttat tggaaaatta
     1261 agggccttgt gcaacccact cgacttcttt tggaatatct tgaagaaaaa tatgaagagc
     1321 atttgtatga gcgcgatgaa ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg
     1381 agtttcccaa tcttccttat tatattgatg gtgatgttaa attaacacag tctatggcca
     1441 tcatacgtta tatagctgac aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag
     1501 agatttcaat gcttgaagga gcggttttgg atattagata cggtgtttcg agaattgcat
     1561 atagtaaaga ctttgaaact ctcaaagttg attttcttag caagctacct gaaatgctga
     1621 aaatgttcga agatcgttta tgtcataaaa catatttaaa tggtgatcat gtaacccatc
     1681 ctgacttcat gttgtatgac gctcttgatg ttgttttata catggaccca atgtgcctgg
     1741 atgcgttccc aaaattagtt tgttttaaaa aacgtattga agctatccca caaattgata
     1801 agtacttgaa atccagcaag tatatagcat ggcctttgca gggctggcaa gccacgtttg
     1861 gtggtggcga ccatcctcca aaatagctcg agtctggtaa agaaaccgct gctgcgaaat
     1921 ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca
     1981 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2041 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     2101 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2161 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2221 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2281 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2341 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2401 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2461 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2521 aacgtttaca atttctggcg gcacgatggc atgagattat caaaaaggat cttcacctag
     2581 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2641 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2701 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2761 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2821 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2881 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2941 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3001 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3061 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3121 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3181 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3241 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3301 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3361 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3421 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3481 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatcatg attgaagcat
     3541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3601 aataggtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3661 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3721 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     3781 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     3841 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3901 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3961 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4021 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4081 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4141 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4201 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4261 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4321 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4381 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4441 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4501 accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     4561 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     4621 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4681 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     4741 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     4801 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     4861 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     4921 ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg ggttactgat
     4981 gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt atggatgcgg
     5041 cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac agatgtaggt
     5101 gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat ggtgcagggc
     5161 gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat tcatgttgtt
     5221 gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg tatcggtgat
     5281 tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa cgacaggagc
     5341 acgatcatgc tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc
     5401 aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg
     5461 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     5521 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     5581 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     5641 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     5701 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg
     5761 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5821 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5881 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5941 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     6001 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     6061 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     6121 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     6181 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     6241 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     6301 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     6361 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     6421 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     6481 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     6541 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     6601 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     6661 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     6721 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6781 cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg
     6841 cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg
     6901 cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact ata
//
