LOCUS       sequence-489445-        2682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription of mutant human FAM136A downstream of SP6 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2682)
  AUTHORS   Harhai M, Foged MM, Zarges C, Landoni JC, Chollet S, Simonelli M, 
            Recazens E, Lisci M, Laban N, Manley S, Riemer J, Lopez-Escamez JA, 
            Lysakowski A, Jourdain AA
  TITLE     An updated inventory of genes essential for oxidative 
            phosphorylation identifies a mitochondrial origin in familial 
            Meniere's disease.
  JOURNAL   Cell Rep. 2025 Jul 25;44(8):116069. doi: 
            10.1016/j.celrep.2025.116069.
  PUBMED    40714634
REFERENCE   2  (bases 1 to 2682)
  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: "Cell Rep.";
            date: "2025-07-25"; volume: "44(8):116069. doi"; pages: " 
            10.1016/j.celrep.2025.116069"
COMMENT     Sequence Label: pTwist Amp High Copy FAM136A-HA mutant
FEATURES             Location/Qualifiers
     source          1..2682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    598..622
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     rep_origin      739..1327
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1228..1247
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1480..1502
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1499..1515
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1577..1593
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1612..1631
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1612..1630
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(1681..1707)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1955..1973)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1956..1973)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1981..1997)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2085..2101)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2215..2319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2320..2388,2389..2682,1..498)
                     /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(2538..2557)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
       61 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
      121 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
      181 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
      241 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
      301 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggctctcgc
      361 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
      421 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
      481 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
      541 aaacttcatt tttaatttaa aaggatctag gtgaagatcc ttgaattgcc attatcaaca
      601 agtttgtaca aaaaagcagg ctccgaattg gtatcacgag gacgatttga agccccctac
      661 ccatttacgc tgacttgacg ggacggcggc tttgttgaat agagcgtcag accccgtaga
      721 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      781 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      841 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      901 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      961 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1021 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1081 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1141 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1201 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1261 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1321 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1381 tcacatgttc tttccagcga catatgccat acgccgaact gaactccacc taatgtcttg
     1441 gagattgtcg gatagcgcgg aataggattg tgccctctga gcggataaca atttcacaca
     1501 ggaaacagct atgaccatga cacatgcatc caccatcgca gacttatcat cactgagcct
     1561 ccacctagcc ttcacacagg aaacagctat gaccatgatt acgccaagct ctaatacgac
     1621 tcactatagg gagacaagct tgcatgcctg caggtcgact ctagaggatc ctcactatta
     1681 agcgtaatct ggaacatcgt atgggtagaa cttctctaat tcacttgtga cgagagcctg
     1741 ggcctgggcc agtggaacat gacatctctc aatgcattga tggacctgtt tcattgaggc
     1801 ctgactgtct tcgcaacagc ttgcagagca tcggaacatc aatccctgca tcttacgtat
     1861 attctcccgc tctaagcttt tcaccatgct ctcgactgcc tcctgaacac gtaactgctg
     1921 caattctgcc atggtaccga gctcgaattc gtattctata gtgtcaccta aatccaattc
     1981 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aaaggtcagg
     2041 cggaatggca cttccttatt ccagatgcgt gcggattatg catgactggc cgtcgtttta
     2101 caacgtcgtg actgggaaaa ccctggcgct tgtggtggat cacagttatt ggagacttcc
     2161 ttggccgtac gatgggtttt gtttcaggta tctggcactt ttcggggaaa tgtgcgcgga
     2221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     2281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     2341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     2401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     2461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     2521 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2641 gaaaagcatc ttacggatgg catgacagta agagaattat gc
//
