LOCUS       sequence-364034-        7703 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of ANXA2-mRuby3 (Annexin A2-mRuby3) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7703)
  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 7703)
  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-ANXA2-mRuby3
FEATURES             Location/Qualifiers
     source          1..7703
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1507..1529
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1545..1561
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1899..1915
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    3839..4405
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3855..4407
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3999..4016)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4226..4245
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4425..4799
                     /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             5029..5498
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5628..5645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5799..6387)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5879..5898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6551..7342,7343..7411))
                     /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     7174..7193
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7412..7516)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7584..7602
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
      601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
      661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
      721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
      841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
      901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
      961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat
     1981 aaagcttaaa ggatccatgg gccgccagct agcggggtgc ggagacgctg ggaagaaggc
     2041 ttccttcaaa atgtctactg ttcacgaaat cctgtgcaag ctcagcttgg agggtgatca
     2101 ctctacaccc ccaagtgcat atgggtctgt caaagcctat actaactttg atgctgagcg
     2161 ggatgctttg aacattgaaa cagccatcaa gaccaaaggt gtggatgagg tcaccattgt
     2221 caacattttg accaaccgca gcaatgcaca gagacaggat attgccttcg cctaccagag
     2281 aaggaccaaa aaggaacttg catcagcact gaagtcagcc ttatctggcc acctggagac
     2341 ggtgattttg ggcctattga agacacctgc tcagtatgac gcttctgagc taaaagcttc
     2401 catgaagggg ctgggaaccg acgaggactc tctcattgag atcatctgct ccagaaccaa
     2461 ccaggagctg caggaaatta acagagtcta caaggaaatg tacaagactg atctggagaa
     2521 ggacattatt tcggacacat ctggtgactt ccgcaagctg atggttgccc tggcaaaggg
     2581 tagaagagca gaggatggct ctgtcattga ttatgaactg attgaccaag atgctcggga
     2641 tctctatgac gctggagtga agaggaaagg aactgatgtt cccaagtgga tcagcatcat
     2701 gaccgagcgg agcgtgcccc acctccagaa agtatttgat aggtacaaga gttacagccc
     2761 ttatgacatg ttggaaagca tcaggaaaga ggttaaagga gacctggaaa atgctttcct
     2821 gaacctggtt cagtgcattc agaacaagcc cctgtatttt gctgatcggc tgtatgactc
     2881 catgaagggc aaggggacgc gagataaggt cctgatcaga atcatggtct cccgcagtga
     2941 agtggacatg ttgaaaatta ggtctgaatt caagagaaag tacggcaagt ccctgtacta
     3001 ttatatccag caagacacta agggcgacta ccagaaagcg ctgctgtacc tgtgtggtgg
     3061 agatgacgga tccgggaatt cgggctctgg ttcaggcgtg agcaagggcg aggagctcat
     3121 caaggagaac atgcggatga aagtcgtcat ggaaggctct gtcaatgggc accagttcaa
     3181 gtgtaccgga gaaggggaag ggcgtcccta tgaaggcgta caaaccatgc gcatcaaagt
     3241 gatagagggc gggccactcc cttttgcctt cgacattctg gctacctcct tcatgtatgg
     3301 cagtcggaca ttcatcaagt atcccgcaga cattccggac ttcttcaaac agtcctttcc
     3361 agaaggcttt acatgggaga gggtgactag gtacgaagat ggcggagtgg ttactgtgac
     3421 tcaggacacg tcattggagg acggagagct ggtgtacaat gtgaaagtac gcggagtgaa
     3481 ctttcccagc aatggtcctg ttatgcagaa gaaaaccaag ggttgggaac ccaatacgga
     3541 gatgatgtat ccagctgatg gtggcctgag gggatacaca gacatagccc tgaaagtcga
     3601 tggaggtggc catctgcact gcaactttgt gaccacctac agaagcaaga aaactgtggg
     3661 gaacattaag atgcctggag tgcatgcagt cgatcacaga cttgagagaa tcgaggagtc
     3721 tgataacgag acatacgtgg tacaacgaga agttgccgtt gcgaagtaca gcaatctggg
     3781 cgggggcatg gacgagctgt acaagtaatg atttctcgag agcggccgct acgtaaattc
     3841 cgcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     3901 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     3961 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     4021 tctgttgaat gtcgtgaagg aagcagttcc tctggaagtt tcttgaagac aaacaacgtc
     4081 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     4141 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     4201 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     4261 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     4321 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     4381 tttcctttga aaaacacgat gataatatgg ccacaaccgt cgacatggcc aagttgacca
     4441 gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc
     4501 ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg
     4561 tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc ctggcctggg
     4621 tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact
     4681 tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg gggcgggagt
     4741 tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag caggactgag
     4801 tcgaccgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatggct
     4861 cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     4921 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     4981 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     5041 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     5101 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     5161 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     5221 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     5281 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     5341 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     5401 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     5461 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta
     5521 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc
     5581 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     5641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6121 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6181 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6241 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6301 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6361 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6421 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6481 ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt
     6541 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6601 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6661 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6721 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6781 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6841 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6901 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6961 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7021 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7081 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7141 gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt
     7201 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7261 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7321 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7381 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7441 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7501 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     7561 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcactata
     7621 ggagcttggc ccattgcata cgttgtatcc atatcataat atgtacattt atattggctc
     7681 atgtccaaca ttaccgccat gtt
//
