LOCUS       sequence-364091-        7670 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of ANXA1-mRuby3 (Annexin A1-mRuby3) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7670)
  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 7670)
  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-ANXA1-mRuby3
FEATURES             Location/Qualifiers
     source          1..7670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        23..402
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    761..1118
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1183..1599
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1529..1551
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1567..1583
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1609..1983
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1921..1937
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             complement(2136..2147)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     misc_feature    3828..4394
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3844..4396
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3988..4005)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4215..4234
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4414..4788
                     /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             5018..5487
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5617..5634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5788..6376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5868..5887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6540..7331,7332..7400))
                     /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     7163..7182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7401..7505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7573..7591
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
       61 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      121 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      181 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      241 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      301 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      361 acttggcagt acatctacgt attagtcatc gctattacca tgctagagaa ccatcagatg
      421 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
      481 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
      541 cccctcactc ggcgcgccag tcctccgatt gactgagtcg cccgggtacc cgtgtatcca
      601 ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct
      661 ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc cgggatcggg
      721 agacccctgc ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc
      781 tgtgtctgtc cgattgtcta gtgtctatga ctgattttat gcgcctgcgt cggtactagt
      841 tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc ggaacacccg
      901 gccgcaaccc tgggagacgt cccagggact tcgggggccg tttttgtggc ccgacctgag
      961 tccaaaaatc ccgatcgttt tggactcttt ggtgcacccc ccttagagga gggatatgtg
     1021 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg
     1081 gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca gcatcgttct gtgttgtctc
     1141 tgtctgactg tgtttctgta tttgtctgaa aatatgggcc cgggccagac tgttaccact
     1201 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
     1261 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
     1321 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
     1381 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
     1441 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
     1501 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
     1561 tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc atatgagatc
     1621 ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac aagagttact
     1681 aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca cgaagtctgg
     1741 agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc tcacccttac
     1801 cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga acctcgctgg
     1861 aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga cggcatcgca
     1921 gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc atcctctaga
     1981 ctgccggatc tgccatcgat ataaagctta aaggatccat ggcaatggta tcagaattcc
     2041 tcaagcaggc ctggtttatt gaaaatgaag agcaggaata tgttcaaact gtgaagtcat
     2101 ccaaaggtgg tcccggatca gcggtgagcc cctatcctac cttcaatcca tcctcggatg
     2161 tcgctgcctt gcataaggcc ataatggtta aaggtgtgga tgaagcaacc atcattgaca
     2221 ttctaactaa gcgaaacaat gcacagcgtc aacagatcaa agcagcatat ctccaggaaa
     2281 caggaaagcc cctggatgaa acactgaaga aagcccttac aggtcacctt gaggaggttg
     2341 ttttagctct gctaaaaact ccagcgcaat ttgatgctga tgaacttcgt gctgccatga
     2401 agggccttgg aactgatgaa gatactctaa ttgagatttt ggcatcaaga actaacaaag
     2461 aaatcagaga cattaacagg gtctacagag aggaactgaa gagagatctg gccaaagaca
     2521 taacctcaga cacatctgga gattttcgga acgctttgct ttctcttgct aagggtgacc
     2581 gatctgagga ctttggtgtg aatgaagact tggctgattc agatgccagg gccttgtatg
     2641 aagcaggaga aaggagaaag gggacagacg taaacgtgtt caataccatc cttaccacca
     2701 gaagctatcc acaacttcgc agagtgtttc agaaatacac caagtacagt aagcatgaca
     2761 tgaacaaagt tctggacctg gagttgaaag gtgacattga gaaatgcctc acagctatcg
     2821 tgaagtgcgc cacaagcaaa ccagctttct ttgcagagaa gcttcatcaa gccatgaaag
     2881 gtgttggaac tcgccataag gcattgatca ggattatggt ttcccgttct gaaattgaca
     2941 tgaatgatat caaagcattc tatcagaaga tgtatggtat ctccctttgc caagccatcc
     3001 tggatgaaac caaaggagat tatgagaaaa tcctggtggc tctttgtgga ggaaacggat
     3061 ccgggaattc gggctctggt tcaggcgtga gcaagggcga ggagctcatc aaggagaaca
     3121 tgcggatgaa agtcgtcatg gaaggctctg tcaatgggca ccagttcaag tgtaccggag
     3181 aaggggaagg gcgtccctat gaaggcgtac aaaccatgcg catcaaagtg atagagggcg
     3241 ggccactccc ttttgccttc gacattctgg ctacctcctt catgtatggc agtcggacat
     3301 tcatcaagta tcccgcagac attccggact tcttcaaaca gtcctttcca gaaggcttta
     3361 catgggagag ggtgactagg tacgaagatg gcggagtggt tactgtgact caggacacgt
     3421 cattggagga cggagagctg gtgtacaatg tgaaagtacg cggagtgaac tttcccagca
     3481 atggtcctgt tatgcagaag aaaaccaagg gttgggaacc caatacggag atgatgtatc
     3541 cagctgatgg tggcctgagg ggatacacag acatagccct gaaagtcgat ggaggtggcc
     3601 atctgcactg caactttgtg accacctaca gaagcaagaa aactgtgggg aacattaaga
     3661 tgcctggagt gcatgcagtc gatcacagac ttgagagaat cgaggagtct gataacgaga
     3721 catacgtggt acaacgagaa gttgccgttg cgaagtacag caatctgggc gggggcatgg
     3781 acgagctgta caagtaatga tttctcgaga gcggccgcta cgtaaattcc gccccccccc
     3841 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat
     3901 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct
     3961 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg
     4021 tcgtgaagga agcagttcct ctggaagttt cttgaagaca aacaacgtct gtagcgaccc
     4081 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg
     4141 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg
     4201 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga
     4261 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt
     4321 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa
     4381 aaacacgatg ataatatggc cacaaccgtc gacatggcca agttgaccag tgccgttccg
     4441 gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg gctcgggttc
     4501 tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt gaccctgttc
     4561 atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt gtgggtgcgc
     4621 ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt ccgggacgcc
     4681 tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt cgccctgcgc
     4741 gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgagt cgaccgcccg
     4801 ccccacgacc cgcagcgccc gaccgaaagg agcgcacgac cccatggctc gacgataaaa
     4861 taaaagattt tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt
     4921 ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata
     4981 gagaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata
     5041 tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat
     5101 gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat
     5161 ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg
     5221 tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct
     5281 cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact
     5341 cggggcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct
     5401 cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
     5461 tgactacccg tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt
     5521 tttcttaagt atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg
     5581 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5641 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5701 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5761 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5821 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5881 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5941 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6001 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6061 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6121 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6181 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6241 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6301 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6361 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6421 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6481 ccttttgcgg ccggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6541 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6601 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6661 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6721 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6781 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6841 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6901 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6961 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7021 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7081 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7141 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7201 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7261 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7321 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7381 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7441 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7501 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7561 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcactatag gagcttggcc
     7621 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca
//
