LOCUS       sequence-424070-        5765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged VDAC1 in mammalian cells. GFP is attached to 
            the N-terminus of VDAC1 with a flexible linker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5765)
  AUTHORS   Pilic J, Oflaz FE, Gottschalk B, Erdogan YC, Graier WF, Malli R
  TITLE     Visualizing interactions of VDAC1 in live cells using a 
            tetracysteine tag
  JOURNAL   bioRxiv 2023.09.08.556841
REFERENCE   2  (bases 1 to 5765)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2023.09.08.556841"
COMMENT     Sequence Label: pTwist GFP-Linker-VDAC1
FEATURES             Location/Qualifiers
     source          1..5765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      257..393
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    545..616
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      complement(621..667)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    685..733
                     /note="synthetic polyadenylation signal"
     enhancer        814..1117
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1119..1321
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1272..1292
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1392..1394,1395..1397,1398..2108)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1437..1458)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1698..1717)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2045..2066
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3025..3146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3036..3055)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3090..3109
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      3261..3307
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3337..3473
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    3518..3589
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3815..4403)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3895..3914)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(5149..5217,5218..5765,1..244)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5367..5386)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
       61 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
      121 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
      181 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
      241 gtaaccgatt ctaggtgcat tggcgcagaa aaaaatgcct gatgcgacgc tgcgcgtctt
      301 atactcccac atatgccaga ttcagcaacg gatacggctt ccccaacttg cccacttcca
      361 tacgtgtcct ccttaccaga aatttatcct taagatcccg aatcgtttaa actcgactct
      421 ggctctatcg aatctccgtc gtttcgagct tacgcgaaca gccgtggcgc tcatttgctc
      481 gtcgggcatc gaatctcgtc agctatcgtc agcttacctt tttggcagcg atcgcggctc
      541 ccgacatctt ggaccattag ctccacaggt atcttcttcc ctctagtggt cataacagca
      601 gcttcagcta cctctcaatt caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt
      661 tatgctatca atcgttgcgt tacacacaca aaaaaccaac acacatccat cttcgatgga
      721 tagcgatttt attatctaac tgctgatcga gtgtagccag atctagtaat caattacggg
      781 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      841 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      901 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      961 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     1021 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     1081 gcagtacatc tacgtattag tcatcgctat taccatgctg atgcggtttt ggcagtacat
     1141 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     1201 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     1261 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     1321 tggtttagtg aaccgtcaga tcagatcttt gtcgatccta ccatccactc gacacacccg
     1381 ccagcggccg catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1441 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1501 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1561 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1621 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1681 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1741 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1801 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1861 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1921 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1981 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2041 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2101 tgtacaaggg tggaggcggt agcatggctg tgccacccac gtatgccgat cttggcaaat
     2161 ctgccaggga tgtcttcacc aagggctatg gatttggctt aataaagctt gatttgaaaa
     2221 caaaatctga gaatggattg gaatttacaa gctcaggctc agccaacact gagaccacca
     2281 aagtgacggg cagtctggaa accaagtaca gatggactga gtacggcctg acgtttacag
     2341 agaaatggaa taccgacaat acactaggca ccgagattac tgtggaagat cagcttgcac
     2401 gtggactgaa gctgaccttc gattcatcct tctcacctaa cactgggaaa aaaaatgcta
     2461 aaatcaagac agggtacaag cgggagcaca ttaacctggg ctgcgacatg gatttcgaca
     2521 ttgctgggcc ttccatccgg ggtgctctgg tgctaggtta cgagggctgg ctggccggct
     2581 accagatgaa ttttgagact gcaaaatccc gagtgaccca gagcaacttt gcagttggct
     2641 acaagactga tgaattccag cttcacacta atgtgaatga cgggacagag tttggcggct
     2701 ccatttacca gaaagtgaac aagaagttgg agaccgctgt caatcttgcc tggacagcag
     2761 gaaacagtaa cacgcgcttc ggaatagcag ccaagtatca gattgaccct gacgcctgct
     2821 tctcggctaa agtgaacaac tccagcctga taggtttagg atacactcag actctaaagc
     2881 caggtattaa actgacactg tcagctcttc tggatggcaa gaacgtcaat gctggtggcc
     2941 acaagcttgg tctaggactg gaatttcaag cataagctag cttgactgac tgagatacag
     3001 cgtaccttca gctcacagac atgataagat acattgatga gtttggacaa accacaacta
     3061 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     3121 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     3181 ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtattg
     3241 gcccatctct atcggtatcg tagcataacc ccttggggcc tctaaacggg tcttgagggg
     3301 ttttttgtgc ccctcgggcc ggattgctat ctaccggcat tggcgcagaa aaaaatgcct
     3361 gatgcgacgc tgcgcgtctt atactcccac atatgccaga ttcagcaacg gatacggctt
     3421 ccccaacttg cccacttcca tacgtgtcct ccttaccaga aatttatcct taaggtcgtc
     3481 agctatcctg caggcgatct ctcgatttcg atcaagacat tcctttaatg gtcttttctg
     3541 gacaccacta ggggtcagaa gtagttcatc aaactttctt ccctccctaa tctcattggt
     3601 taccttgggc tatcgaaact taattaacca gtcaagtcag ctacttggcg agatcgactt
     3661 gtctgggttt cgactacgct cagaattgcg tcagtcaagt tcgatctggt ccttgctatt
     3721 gcacccgttc tccgattacg agtttcattt aaatcatgtg agcaaaaggc cagcaaaagg
     3781 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3841 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3901 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3961 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4021 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4081 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4141 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4201 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     4261 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4321 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4381 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4441 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4501 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4561 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4621 ttcgttcatc catagttgca tttaaatttc cgaactctcc aaggccctcg tcggaaaatc
     4681 ttcaaacctt tcgtccgatc catcttgcag gctacctctc gaacgaacta tcgcaagtct
     4741 cttggccggc cttgcgcctt ggctattgct tggcagcgcc tatcgccagg tattactcca
     4801 atcccgaata tccgagatcg ggatcacccg agagaagttc aacctacatc ctcaatcccg
     4861 atctatccga gatccgagga atatcgaaat cggggcgcgc ctggtgtacc gagaacgatc
     4921 ctctcagtgc gagtctcgac gatccatatc gttgcttggc agtcagccag tcggaatcca
     4981 gcttgggacc caggaagtcc aatcgtcaga tattgtactc aagcctggtc acggcagcgt
     5041 accgatctgt ttaaacctag atattgatag tctgatcggt caacgtataa tcgagtccta
     5101 gcttttgcaa acatctatca agagacagga tcagcaggag gctttcgcat gagtattcaa
     5161 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5221 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcgcg agtgggttac
     5281 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgcttt
     5341 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5401 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtattca
     5461 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5521 ataaccatga gtgataacac tgcggccaac ttacttctga caacgattgg aggaccgaag
     5581 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5641 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     5701 gcaacaacct tgcgtaaact attaactggc gaactactta ctctagcttc ccggcaacag
     5761 ttgat
//
