LOCUS       Exported                6226 bp ds-DNA     circular SYN 07-DEC-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6226)
  AUTHORS   Voeltz Lab
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Dec 1, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6226
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    181..250
                     /label=CMV Promoter
     primer_bind     complement(265..307)
                     /label=TN509
     primer_bind     265..307
                     /label=TN506
     misc_feature    289..486
                     /label=TOMM70NTD
                     /note="According to Suzuki H, et al JCS 2002, In rats, 
                     TOMM70 AA's 1-66 are sufficient to localize to mito. 1-69 
                     includes 6 arginines (R's) which is also sufficient to 
                     localize to mito. To ensure localization to mito, will 
                     clone 1-66 in human for fusion protein of 
                     Tom70NTD-ABHD16Acyto."
     CDS             403..465
                     /codon_start=1
                     /label=TOMM70TMD
                     /translation="WQLALAVGAPLLLGAGAIYLW"
     misc_feature    412
                     /label=G > A, Ala to Thr
     primer_bind     474..502
                     /label=TN508
     primer_bind     complement(474..502)
                     /label=TN507
     misc_feature    486..1812
                     /label=ABHD16Acyto
                     /note="no ATG start"
     misc_feature    496..888
                     /label=unknown structural domain
                     /note="Singh S. et al 2020 biochemistry"
     misc_feature    664..666
                     /label=TCC > GCC, S to A
     misc_feature    703..705
                     /label=CAC>GCC, H to A
     misc_feature    718..720
                     /label=GAC>AAC, D to N
     misc_feature    976..1728
                     /label=ABHD domain
     misc_feature    1054..1056
                     /label=TCA > GCA, S to A
     misc_feature    1164
                     /label=A > G
                     /note="wobble"
     misc_feature    1201..1203
                     /label=TCC>GCC S to A
     misc_feature    1426..1428
                     /label=D to N
     misc_feature    1657..1659
                     /label=H to A
     misc_feature    1665
                     /label=C > T
                     /note="wobble mutation"
     misc_feature    1837..1842
                     /label=Kozak
     CDS             1843..2553
                     /label=mNeonGreen [3']
     rep_origin      2813..3253
                     /direction=RIGHT
                     /label=F1 ori
     rep_origin      2813..3253
                     /direction=RIGHT
                     /label=F1 ori
     promoter        3320..3348
                     /label=Amp prom
     promoter        3320..3348
                     /label=Amp prom
     promoter        3552..3748
                     /label=SV40 early promoter
     promoter        3552..3748
                     /label=SV40 early promoter
     misc_feature    3599..3676
                     /label=SV40 ORI
     misc_feature    3599..3676
                     /label=SV40 ORI
     CDS             3786..4577
                     /label=Kan/neoR
     rep_origin      5163..5791
                     /direction=RIGHT
                     /label=ColE1 origin
     rep_origin      5163..5791
                     /direction=RIGHT
                     /label=ColE1 origin
ORIGIN
        1 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
       61 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      121 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      181 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      241 ccgtcagatc cgctagcgct accggactca gatctcgagc tcaagcttat ggccgcctct
      301 aaacctgtgg aggcagcggt ggtcgcagcc gctgtaccga gctccgggag tggggtgggc
      361 ggcggcggga ctgcgggccc gggcacgggg gggctgccgc gatggcagct ggctctggcg
      421 gtcggggcac ccctgctgct gggcgcgggt gccatatacc tgtggagtcg gcagcaacgg
      481 cgccggggcc gctggaccaa cccccagtac cggcagttca tcaccatctt ggaagcaaca
      541 catcggaacc agtcttcaga aaacaagagg cagcttgcca actacaactt tgacttccgg
      601 agctggccag tcgacttcca ctgggaagaa cccagcagcc ggaaggagtc tcgagggggc
      661 ccttcccgcc ggggtgtggc cctgcttcgc ccagagcccc tgcaccgggg gacagcagac
      721 accctcctca accgggttaa gaagctgcct tgtcagatca ccagctacct ggtggcgcac
      781 accctagggc gccggatgct gtatccaggc tctgtgtacc tgctgcagaa ggccctcatg
      841 cctgtgctgc tgcagggcca ggcccgactg gtggaagagt gtaatgggcg ccgggcaaag
      901 ctgctggcct gtgatggcaa tgagattgac accatgtttg tggaccggcg ggggacagct
      961 gagccccagg gacagaagct ggtgatctgc tgtgagggga atgctgggtt ttatgaggtg
     1021 ggctgcgtct ccacgcccct ggaagctgga tattcagtcc tgggctggaa tcatccaggc
     1081 tttgctggaa gcacgggggt gccattcccg cagaatgagg ctaatgccat ggatgtggtg
     1141 gtccagtttg ccatccaccg cctaggcttc cagccccagg acatcatcat ctacgcctgg
     1201 tccatcggcg gcttcactgc cacgtgggca gccatgtcct acccagatgt tagtgccatg
     1261 atcctggatg cctcctttga tgacctggtg cccttggcct tgaaggtcat gccagacagc
     1321 tggaggggcc tggtgaccag gaccgtgagg cagcatctca atctaaacaa cgcggagcag
     1381 ctgtgcagat accagggtcc tgtactgctg atccggagaa ccaaggatga gatcatcacc
     1441 accacggttc ctgaggacat catgtccaac cgaggcaatg acctcctgct gaagctcctg
     1501 cagcatcggt atccccgggt gatggcagag gagggtcttc gagtggtgag gcagtggttg
     1561 gaggcctcct cacagctgga ggaagcctca atttatagcc gatgggaggt ggaagaggac
     1621 tggtgtctgt ctgtcctccg ctcctaccag gcagaacacg ggcccgactt cccctggagc
     1681 gtgggggagg acatgagtgc agatggacgg cggcagctgg ctttgtttct ggctcggaag
     1741 catctgcaca actttgaggc cactcactgc accccactcc cagcccagaa cttccagatg
     1801 ccctggcacc tcccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
     1861 gaggataaca tggcctctct cccagcgaca catgagttac acatctttgg ctccatcaac
     1921 ggtgtggact ttgacatggt gggtcagggc accggcaatc caaatgatgg ttatgaggag
     1981 ttaaacctga agtccaccaa gggtgacctc cagttctccc cctggattct ggtccctcat
     2041 atcgggtatg gcttccatca gtacctgccc taccctgacg ggatgtcgcc tttccaggcc
     2101 gccatggtag atggctccgg ctaccaagtc catcgcacaa tgcagtttga agatggtgcc
     2161 tcccttactg ttaactaccg ctacacctac gagggaagcc acatcaaagg agaggcccag
     2221 gtgaagggga ctggtttccc tgctgacggt cctgtgatga ccaactcgct gaccgctgcg
     2281 gactggtgca ggtcgaagaa gacttacccc aacgacaaaa ccatcatcag tacctttaag
     2341 tggagttaca ccactggaaa tggcaagcgc taccggagca ctgcgcggac cacctacacc
     2401 tttgccaagc caatggcggc taactatctg aagaaccagc cgatgtacgt gttccgtaag
     2461 acggagctca agcactccaa gaccgagctc aacttcaagg agtggcaaaa ggcctttacc
     2521 gatgtgatgg gcatggacga gctgtacaag taagcggccg cgactctaga tcataatcag
     2581 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     2641 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     2701 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2761 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     2821 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     2881 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     2941 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3001 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3061 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3121 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3181 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3241 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3301 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3361 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3421 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3481 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3541 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3601 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3661 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3721 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     3781 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     3841 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     3901 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     3961 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4021 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4081 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4141 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4201 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4261 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4321 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4381 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4441 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4501 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     4561 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     4621 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4681 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4741 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     4801 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     4861 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     4921 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     4981 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5041 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5101 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5161 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5221 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5281 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     5341 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5401 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5461 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5521 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5581 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5641 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5701 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5761 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5821 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5881 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     5941 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6001 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6061 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6121 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6181 cccagtacat gaccttatgg gactttccta cttggcagta catcta
//
