LOCUS       sequence-364037-        7592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of ANXA5-mRuby3 (Annexin A5-mRuby3) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7592)
  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 7592)
  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-ANXA5-mRuby3
FEATURES             Location/Qualifiers
     source          1..7592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    722..1079
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1144..1560
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1490..1512
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1528..1544
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1570..1944
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1882..1898
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     misc_feature    3711..4277
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3727..4279
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3871..3888)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4098..4117
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4297..4671
                     /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             4901..5370
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5500..5517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5671..6259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5751..5770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6423..7214,7215..7283))
                     /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     7046..7065
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7284..7388)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7456..7474
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7576..7592,1..363)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
       61 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      121 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      181 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      241 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      301 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      361 atgctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      421 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      481 gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat tgactgagtc
      541 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      601 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      661 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      721 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      781 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      841 ctgacgagtt cggaacaccc ggccgcaacc ctgggagacg tcccagggac ttcgggggcc
      901 gtttttgtgg cccgacctga gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc
      961 cccttagagg agggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     1021 ccgtctgaat ttttgctttc ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc
     1081 agcatcgttc tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaatatgggc
     1141 ccgggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag
     1201 cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct
     1261 gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc
     1321 atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc
     1381 ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt
     1441 gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct
     1501 cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc
     1561 cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa cttccctgac
     1621 cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca ggctctctac
     1681 ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca actggaccga
     1741 ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg acaccagact
     1801 aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac ccccaccgcc
     1861 ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc tgccgacccc
     1921 gggggtggac catcctctag actgccggat ctgccatcga tataaagctt atggcacagg
     1981 ttctcagagg cactgtgact gacttccctg gatttgatga gcgggctgat gcagaaactc
     2041 ttcggaaggc tatgaaaggc ttgggcacag atgaggagag catcctgact ctgttgacat
     2101 cccgaagtaa tgctcagcgc caggaaatct ctgcagcttt taagactctg tttggcaggg
     2161 atcttctgga tgacctgaaa tcagaactaa ctggaaaatt tgaaaaatta attgtggctc
     2221 tgatgaaacc ctctcggctt tatgatgctt atgaactgaa acatgccttg aagggagctg
     2281 gaacaaatga aaaagtactg acagaaatta ttgcttcaag gacacctgaa gaactgagag
     2341 ccatcaaaca agtttatgaa gaagaatatg gctcaagcct ggaagatgac gtggtggggg
     2401 acacttcagg gtactaccag cggatgttgg tggttctcct tcaggctaac agagaccctg
     2461 atgctggaat tgatgaagct caagttgaac aagatgctca ggctttattt caggctggag
     2521 aacttaaatg ggggacagat gaagaaaagt ttatcaccat ctttggaaca cgaagtgtgt
     2581 ctcatttgag aaaggtgttt gacaagtaca tgactatatc aggatttcaa attgaggaaa
     2641 ccattgaccg cgagacttct ggcaatttag agcaactact ccttgctgtt gtgaaatcta
     2701 ttcgaagtat acctgcctac cttgcagaga ccctctatta tgctatgaag ggagctggga
     2761 cagatgatca taccctcatc agagtcatgg tttccaggag tgagattgat ctgtttaaca
     2821 tcaggaagga gtttaggaag aattttgcca cctctcttta ttccatgatt aagggagata
     2881 catctgggga ctataagaaa gctcttctgc tgctctgtgg agaagatgac aagcttaaag
     2941 gatccgggaa ttcgggctct ggttcaggcg tgagcaaggg cgaggagctc atcaaggaga
     3001 acatgcggat gaaagtcgtc atggaaggct ctgtcaatgg gcaccagttc aagtgtaccg
     3061 gagaagggga agggcgtccc tatgaaggcg tacaaaccat gcgcatcaaa gtgatagagg
     3121 gcgggccact cccttttgcc ttcgacattc tggctacctc cttcatgtat ggcagtcgga
     3181 cattcatcaa gtatcccgca gacattccgg acttcttcaa acagtccttt ccagaaggct
     3241 ttacatggga gagggtgact aggtacgaag atggcggagt ggttactgtg actcaggaca
     3301 cgtcattgga ggacggagag ctggtgtaca atgtgaaagt acgcggagtg aactttccca
     3361 gcaatggtcc tgttatgcag aagaaaacca agggttggga acccaatacg gagatgatgt
     3421 atccagctga tggtggcctg aggggataca cagacatagc cctgaaagtc gatggaggtg
     3481 gccatctgca ctgcaacttt gtgaccacct acagaagcaa gaaaactgtg gggaacatta
     3541 agatgcctgg agtgcatgca gtcgatcaca gacttgagag aatcgaggag tctgataacg
     3601 agacatacgt ggtacaacga gaagttgccg ttgcgaagta cagcaatctg ggcgggggca
     3661 tggacgagct gtacaagtaa tgatttctcg agagcggccg ctacgtaaat tccgcccccc
     3721 cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt
     3781 tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct
     3841 tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga
     3901 atgtcgtgaa ggaagcagtt cctctggaag tttcttgaag acaaacaacg tctgtagcga
     3961 ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac
     4021 gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag
     4081 ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc
     4141 agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg
     4201 tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt
     4261 gaaaaacacg atgataatat ggccacaacc gtcgacatgg ccaagttgac cagtgccgtt
     4321 ccggtgctca ccgcgcgcga cgtcgccgga gcggtcgagt tctggaccga ccggctcggg
     4381 ttctcccggg acttcgtgga ggacgacttc gccggtgtgg tccgggacga cgtgaccctg
     4441 ttcatcagcg cggtccagga ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg
     4501 cgcggcctgg acgagctgta cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac
     4561 gcctccgggc cggccatgac cgagatcggc gagcagccgt gggggcggga gttcgccctg
     4621 cgcgacccgg ccggcaactg cgtgcacttc gtggccgagg agcaggactg agtcgaccgc
     4681 ccgccccacg acccgcagcg cccgaccgaa aggagcgcac gaccccatgg ctcgacgata
     4741 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     4801 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac ataactgaga
     4861 atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg
     4921 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa
     4981 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     5041 gatggtcccc agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca
     5101 gggtgcccca aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct
     5161 tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc
     5221 actcggggcg ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac
     5281 cctcttgcag ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt
     5341 gattgactac ccgtcagcgg gggtctttca catgcagcat gtatcaaaat taatttggtt
     5401 ttttttctta agtatttaca ttaaatggcc atagttgcat taatgaatcg gccaacgcgc
     5461 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5521 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5581 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5641 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5701 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5761 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5821 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5881 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5941 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6001 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6061 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6121 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6181 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6241 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6301 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6361 gatccttttg cggccggccg caaatcaatc taaagtatat atgagtaaac ttggtctgac
     6421 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     6481 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     6541 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     6601 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     6661 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     6721 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     6781 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6841 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6901 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6961 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7021 tgctcttgcc cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg
     7081 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7141 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7201 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     7261 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     7321 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7381 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     7441 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcacta taggagcttg
     7501 gcccattgca tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa
     7561 cattaccgcc atgttgacat tgattattga ct
//
