LOCUS       sequence-469823-        6097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of yeast OMP25.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6097)
  AUTHORS   Zheng C, Nguyen KK, Vishnivetskiy SA, Gurevich VV, Gurevich EV
  TITLE     Arrestin-3 binds parkin and enhances parkin-dependent mitophagy.
  JOURNAL   J Neurochem. 2025 Jan;169(1):e16043. doi: 10.1111/jnc.16043. Epub 
            2024 Jan 9.
  PUBMED    38196269
REFERENCE   2  (bases 1 to 6097)
  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: "J 
            Neurochem."; date: "2025-01"; volume: "169(1):e16043. doi"; pages: "
            10.1111/jnc.16043. Epub 2024 Jan 9"
COMMENT     Sequence Label: pHALgMT
FEATURES             Location/Qualifiers
     source          1..6097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      385..973
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     874..893
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1127..1144
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1261..1282
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1297..1302,1303..1320,1321..1327)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1335..1351
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1340..1362
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1359..1375
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1359..1375
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    1424..1545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1435..1454)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1489..1508
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1719..2513)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1830..1849)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2440..2459
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(2580..2909)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2594..2729
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2648..2667)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     2894..2914
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(2923..3351)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3033..3054)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3245..3264
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    3397..3621
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3610..3627
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        3647..3665
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3647..3664
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(4301..4327)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4340..4366)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(4379..4405)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4433..4452)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4434..4452)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4497..4700)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4526..4546)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4701..5080
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5252..5271
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5346..5450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5451..5519,5520..6097,1..214)
                     /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(5669..5688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
       61 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      121 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      181 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      241 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
      301 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
      361 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
      421 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
      481 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
      541 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
      601 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
      661 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
      721 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
      781 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
      841 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
      901 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
      961 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1021 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1081 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1141 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1201 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1261 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     1321 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     1381 ttacgccaag ctctagctag aggtcgacgg tatacagaca tgataagata cattgatgag
     1441 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     1501 gctattgctt tatttgtaac cattataagc tgcaataaac aagttggggt gggcgaagaa
     1561 ctccagcatg agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc
     1621 gaagcccaac ctttcataga aggcggcggt ggaatcgaaa tctcgtgatg gcaggttggg
     1681 cgtcgcttgg tcggtcattt cgaaccccag agtcccgctc agaagaactc gtcaagaagg
     1741 cgatagaagg cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg
     1801 tcagcccatt cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga
     1861 tagcggtccg ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc
     1921 accatgatat tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc
     1981 atgcgcgcct tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc
     2041 agatcatcct gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt
     2101 ttcgcttggt ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca
     2161 tcagccatga tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc
     2221 ggcacttcgc ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct
     2281 gcgcaaggaa cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ctgcagttca
     2341 ttcagggcac cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc
     2401 cggaacacgg cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc
     2461 ctctccaccc aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac
     2521 gatcctcatc ctgtctcttg atcagatccg aaaatggata tacaagctcc cgggagcttt
     2581 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
     2641 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
     2701 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
     2761 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
     2821 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     2881 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
     2941 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3001 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3061 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3121 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3181 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3241 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3301 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tggggatacc
     3361 ccctagagcc ccagctggtt ctttccgcct cagaagccat agagcccacc gcatccccag
     3421 catgcctgct attgtcttcc caatcctccc ccttgctgtc ctgccccacc ccacccccca
     3481 gaatagaatg acacctactc agacaatgcg atgcaatttc ctcattttat taggaaagga
     3541 cagtgggagt ggcaccttcc agggtcaagg aaggcacggg ggaggggcaa acaacagatg
     3601 gctggcaact agaaggcaca gtcgaggctg atcagcgagc tctagcattt aggtgacact
     3661 atagaatagg gccaagcttt catcagagct gctttcggta tctcacgaag gcccaaactg
     3721 ctaccagggt aagggcaaac actggcagca gcaccacagc tacaggaact ccactaggct
     3781 ctccgtcgcc tcgatgccgc ggcttaagac tgttgatggt tactcggaac agcatggagc
     3841 cgtcgggggt gatcaggcgc tcgtcgataa ttttgttgcc gttccacagg gtccctgtta
     3901 cagtgatctt tttgccgtcg aacacggcga tgccttcata cggccgtccg aaatagttca
     3961 gcatgttcgg cgtaaccccg tcgattacca gtgtgccata gggcaggatc accttaaagt
     4021 gatgatcatc cacagggtac accaccttaa acacctcttc gatctgggcc atttggtcgg
     4081 cgctcagacc ttcatacggg atgatgacat ggatgtcgat cttcagggca ttttcaccgc
     4141 tccggacaat cctttggatc ggagttacgg acacggcgag attctgcagc aaactggaca
     4201 cacctccctg ttcaaggact tggtccaggt tgtaggcggc tgtctgttcc cagtccccaa
     4261 cgaaatcttc gagtgtgaag accatgcttg cggccgctcc agcataatca ggcacatcat
     4321 aagggtatcc ggcgctgcca gcgtagtcag gcacatcata ggggtaggat cctgtgccgg
     4381 cgtaatcagg cacgtcatag ggatacatgg tggtgaattc cagcttgggt ctccctatag
     4441 tgagtcgtat taatttcgat aagccagtaa gcagtgggtt ctctagttag ccagagagct
     4501 ctgcttatat agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag
     4561 ttgttacgac attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg
     4621 tcaatggggt ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg
     4681 tactgccaaa accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa
     4741 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     4801 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     4861 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     4921 ggaacatacg tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg
     4981 ccatttaccg taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc
     5041 gtaattgatt actattaata actagtcaat aatcaatgtc aacgcgtata tctggcccgt
     5101 acatcgcgaa gcagcgcaaa acgcctaacc ctaagcagat tcttcatgca attgtcggtc
     5161 aagccttgcc ttgttgtagc ttaaattttg ctcgcgcact actcagcgac ctccaacaca
     5221 caagcaggga gcagatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     5281 caccataggg gatcgggaga tctcccgatc cgtcgacgtc aggtggcact tttcggggaa
     5341 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5401 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5461 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5521 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5581 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5641 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5701 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5761 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5821 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5881 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5941 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6001 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6061 aattaataga ctggatggag gcggataaag ttgcagg
//
