LOCUS       sequence-364035-        8651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of ANXA6-mRuby3 (Annexin A6-mRuby3) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8651)
  AUTHORS   Yim WW, Yamamoto H, Mizushima N
  TITLE     Annexins A1 and A2 are recruited to larger lysosomal injuries 
            independently of ESCRTs to promote repair.
  JOURNAL   FEBS Lett. 2022 Mar 11. doi: 10.1002/1873-3468.14329.
  PUBMED    35274304
REFERENCE   2  (bases 1 to 8651)
  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: "FEBS Lett. 
            2022 Mar 11. doi: 10.1002/1873-3468.14329."
COMMENT     Sequence Label: pMRX-IZ-ANXA6-mRuby3
FEATURES             Location/Qualifiers
     source          1..8651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        29..408
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    767..1124
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1189..1605
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1535..1557
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1573..1589
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1615..1989
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1927..1943
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    4815..5381
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4831..5383
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4975..4992)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5202..5221
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5401..5775
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     LTR             6005..6474
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6604..6621)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6775..7363)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6855..6874)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7527..8318,8319..8387))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8150..8169
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8388..8492)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8560..8578
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ggctcatgtc caacattacc gccatgttga cattgattat tgactagtta ttaatagtaa
       61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct agagaaccat
      421 cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt tgaactaacc
      481 aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca ataaaagagc
      541 ccacaacccc tcactcggcg cgccagtcct ccgattgact gagtcgcccg ggtacccgtg
      601 tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc cttgggaggg
      661 tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg ctcgtccggg
      721 atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc tggccagcaa
      781 cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt
      841 actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac gagttcggaa
      901 cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt tgtggcccga
      961 cctgagtcca aaaatcccga tcgttttgga ctctttggtg cacccccctt agaggaggga
     1021 tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc tgaatttttg
     1081 ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt
     1141 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata tgggcccggg ccagactgtt
     1201 accactccct taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac
     1261 cagtcggtag atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc
     1321 tttaacgtcg gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag
     1381 atcaaggtct tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc
     1441 tgggaagcct tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct
     1501 ccgcctcctc ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg
     1561 cctcgatcct ccctttatcc agccctcact ccttctctag gcgcccccat atggccatat
     1621 gagatcttat atggggcacc cccgcccctt gtaaacttcc ctgaccctga catgacaaga
     1681 gttactaaca gcccctctct ccaagctcac ttacaggctc tctacttagt ccagcacgaa
     1741 gtctggagac ctctggcggc agcctaccaa gaacaactgg accgaccggt ggtacctcac
     1801 ccttaccgag tcggcgacac agtgtgggtc cgccgacacc agactaagaa cctagaacct
     1861 cgctggaaag gaccttacac agtcctgctg accaccccca ccgccctcaa agtagacggc
     1921 atcgcagctt ggatacacgc cgcccacgtg aaggctgccg accccggggg tggaccatcc
     1981 tctagactgc cggatctgcc atcgatataa agcttatggc caaaccagca cagggtgcca
     2041 agtaccgggg ctccatccat gacttcccag gctttgaccc caaccaggat gccgaggctc
     2101 tgtacactgc catgaagggc tttggcagtg acaaggaggc catactggac ataatcacct
     2161 cacggagcaa caggcagagg caggaggtct gccagagcta caagtccctc tacggcaagg
     2221 acctcattgc tgatttaaag tatgaattga cgggcaagtt tgaacggttg attgtgggcc
     2281 tgatgaggcc acctgcctat tgtgatgcca aagaaattaa agatgccatc tcgggcattg
     2341 gcactgatga gaagtgcctc attgagatct tggcttcccg gaccaatgag cagatgcacc
     2401 agctggtggc agcatacaaa gatgcctacg agcgggacct ggaggctgac atcatcggcg
     2461 acacctctgg ccacttccag aagatgcttg tggtcctgct ccagggaacc agggaggagg
     2521 atgacgtagt gagcgaggac ctggtacaac aggatgtcca ggacctatac gaggcagggg
     2581 aactgaaatg gggaacagat gaagcccagt tcatttacat cttgggaaat cgcagcaagc
     2641 agcatcttcg gttggtgttc gatgagtatc tgaagaccac agggaagccg attgaagcca
     2701 gcatccgagg ggagctgtct ggggactttg agaagctaat gctggccgta gtgaagtgta
     2761 tccggagcac cccggaatat tttgctgaaa ggctcttcaa ggctatgaag ggcctgggga
     2821 ctcgggacaa caccctgatc cgcatcatgg tctcccgtag tgagttggac atgctcgaca
     2881 ttcgggagat cttccggacc aagtatgaga agtccctcta cagcatgatc aagaatgaca
     2941 cctctggcga gtacaagaag actctgctga agctgtctgg gggagatgat gatgctgctg
     3001 gccagttctt cccggaggca gcgcaggtgg cctatcagat gtgggaactt agtgcagtgg
     3061 cccgagtaga gctgaaggga actgtgcgcc cagccaatga cttcaaccct gacgcagatg
     3121 ccaaagcgct gcggaaagcc atgaagggac tcgggactga cgaagacaca atcatcgata
     3181 tcatcacgca ccgcagcaat gtccagcggc agcagatccg gcagaccttc aagtctcact
     3241 ttggccggga cttaatgact gacctgaagt ctgagatctc tggagacctg gcaaggctga
     3301 ttctggggct catgatgcca ccggcccatt acgatgccaa gcagttgaag aaggccatgg
     3361 agggagccgg cacagatgaa aaggctctta ttgaaatcct ggccactcgg accaatgctg
     3421 aaatccgggc catcaatgag gcctataagg aggactatca caagtccctg gaggatgctc
     3481 tgagctcaga cacatctggc cacttcagga ggatcctcat ttctctggcc acggggcatc
     3541 gtgaggaggg aggagaaaac ctggaccagg cacgggaaga tgcccaggtg gctgctgaga
     3601 tcttggaaat agcagacaca cccagtggag acaaaacttc cttggagaca cgtttcatga
     3661 cgatcctgtg tacccggagc tatccgcacc tccggagagt cttccaggag ttcatcaaga
     3721 tgaccaacta tgacgtggag cacaccatca agaaggagat gtctggggat gtcaaggatg
     3781 catttgtggc cattgttcaa agtgtcaaga acaagcctct cttctttgcc gacaaacttt
     3841 acaaatccat gaagggtgct ggcacagatg agaagactct gaccaggatc atggtatccc
     3901 gcagtgagat tgacctgctc aacatccgga gggaattcat tgagaaatat gacaagtctc
     3961 tccaccaagc cattgagggt gacacctccg gagacttcct gaaggccttg ctggctctct
     4021 gtggtggtga ggacaagctt aaaggatccg ggaattcggg ctctggttca ggcgtgagca
     4081 agggcgagga gctcatcaag gagaacatgc ggatgaaagt cgtcatggaa ggctctgtca
     4141 atgggcacca gttcaagtgt accggagaag gggaagggcg tccctatgaa ggcgtacaaa
     4201 ccatgcgcat caaagtgata gagggcgggc cactcccttt tgccttcgac attctggcta
     4261 cctccttcat gtatggcagt cggacattca tcaagtatcc cgcagacatt ccggacttct
     4321 tcaaacagtc ctttccagaa ggctttacat gggagagggt gactaggtac gaagatggcg
     4381 gagtggttac tgtgactcag gacacgtcat tggaggacgg agagctggtg tacaatgtga
     4441 aagtacgcgg agtgaacttt cccagcaatg gtcctgttat gcagaagaaa accaagggtt
     4501 gggaacccaa tacggagatg atgtatccag ctgatggtgg cctgagggga tacacagaca
     4561 tagccctgaa agtcgatgga ggtggccatc tgcactgcaa ctttgtgacc acctacagaa
     4621 gcaagaaaac tgtggggaac attaagatgc ctggagtgca tgcagtcgat cacagacttg
     4681 agagaatcga ggagtctgat aacgagacat acgtggtaca acgagaagtt gccgttgcga
     4741 agtacagcaa tctgggcggg ggcatggacg agctgtacaa gtaatgattt ctcgagagcg
     4801 gccgctacgt aaattccgcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4861 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4921 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     4981 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagtttctt
     5041 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5101 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5161 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5221 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5281 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5341 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccgtcgac
     5401 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
     5461 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
     5521 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
     5581 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
     5641 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
     5701 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
     5761 gaggagcagg actgagtcga ccgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     5821 gcacgacccc atggctcgac gataaaataa aagattttat ttagtctcca gaaaaagggg
     5881 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     5941 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     6001 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     6061 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     6121 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     6181 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     6241 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     6301 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     6361 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     6421 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     6481 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagtt
     6541 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     6601 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     6661 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     6721 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     6781 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     6841 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     6901 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     6961 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     7021 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     7081 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     7141 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     7201 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     7261 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     7321 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     7381 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     7441 attatcaaaa aggatcttca cctagatcct tttgcggccg gccgcaaatc aatctaaagt
     7501 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7561 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     7621 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     7681 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     7741 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     7801 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     7861 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     7921 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7981 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8041 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8101 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg
     8161 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8221 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8281 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8341 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8401 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     8461 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     8521 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     8581 tttcgtcttc actataggag cttggcccat tgcatacgtt gtatccatat cataatatgt
     8641 acatttatat t
//
