LOCUS       sequence-471672-        7568 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of ANXA4-mRuby3 (Annexin A4-mRuby3) in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7568)
  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 7568)
  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-ANXA4-mRuby3
FEATURES             Location/Qualifiers
     source          1..7568
                     /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    3704..4270
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3720..4272
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3864..3881)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4091..4110
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4290..4664
                     /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             4894..5363
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5493..5510)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5664..6252)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5744..5763)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6416..7207,7208..7276))
                     /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     7039..7058
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7277..7381)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7449..7467
                     /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 gccggatcca tggccatggc
     1981 aaccaaagga ggtactgtca aagctgcttc aggattcaat gccatggaag atgcccagac
     2041 cctgaggaag gccatgaaag ggctcggcac cgatgaagac gccattatta gcgtccttgc
     2101 ctaccgcaac accgcccagc gccaggagat caggacagcc tacaagagca ccatcggcag
     2161 ggacttgata gacgacctga agtcagaact gagtggcaac ttcgagcagg tgattgtggg
     2221 gatgatgacg cccacggtgc tgtatgacgt gcaagagctg cgaagggcca tgaagggagc
     2281 cggcactgat gagggctgcc taattgagat cctggcctcc cggacccctg aggagatccg
     2341 gcgcataagc caaacctacc agcagcaata tggacggagc cttgaagatg acattcgctc
     2401 tgacacatcg ttcatgttcc agcgagtgct ggtgtctctg tcagctggtg ggagggatga
     2461 aggaaattat ctggacgatg ctctcgtgag acaggatgcc caggacctgt atgaggctgg
     2521 agagaagaaa tgggggacag atgaggtgaa atttctaact gttctctgtt cccggaaccg
     2581 aaatcacctg ttgcatgtgt ttgatgaata caaaaggata tcacagaagg atattgaaca
     2641 gagtattaaa tctgaaacat ctggtagctt tgaagatgct ctgctggcta tagtaaagtg
     2701 catgaggaac aaatctgcat attttgctga aaagctctat aaatcgatga agggcttggg
     2761 caccgatgat aacaccctca tcagagtgat ggtttctcga gcagaaattg acatgttgga
     2821 tatccgggca cacttcaaga gactctatgg aaagtctctg tactcgttca tcaagggtga
     2881 cacatctgga gactacagga aagtactgct tgttctctgt ggaggagatg atggatccgg
     2941 gaattcgggc tctggttcag gcgtgagcaa gggcgaggag ctcatcaagg agaacatgcg
     3001 gatgaaagtc gtcatggaag gctctgtcaa tgggcaccag ttcaagtgta ccggagaagg
     3061 ggaagggcgt ccctatgaag gcgtacaaac catgcgcatc aaagtgatag agggcgggcc
     3121 actccctttt gccttcgaca ttctggctac ctccttcatg tatggcagtc ggacattcat
     3181 caagtatccc gcagacattc cggacttctt caaacagtcc tttccagaag gctttacatg
     3241 ggagagggtg actaggtacg aagatggcgg agtggttact gtgactcagg acacgtcatt
     3301 ggaggacgga gagctggtgt acaatgtgaa agtacgcgga gtgaactttc ccagcaatgg
     3361 tcctgttatg cagaagaaaa ccaagggttg ggaacccaat acggagatga tgtatccagc
     3421 tgatggtggc ctgaggggat acacagacat agccctgaaa gtcgatggag gtggccatct
     3481 gcactgcaac tttgtgacca cctacagaag caagaaaact gtggggaaca ttaagatgcc
     3541 tggagtgcat gcagtcgatc acagacttga gagaatcgag gagtctgata acgagacata
     3601 cgtggtacaa cgagaagttg ccgttgcgaa gtacagcaat ctgggcgggg gcatggacga
     3661 gctgtacaag taatgatttc tcgagagcgg ccgctacgta aattccgccc ccccccctaa
     3721 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     3781 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     3841 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     3901 gaaggaagca gttcctctgg aagtttcttg aagacaaaca acgtctgtag cgaccctttg
     3961 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     4021 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     4081 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     4141 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     4201 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     4261 acgatgataa tatggccaca accgtcgaca tggccaagtt gaccagtgcc gttccggtgc
     4321 tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc
     4381 gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca
     4441 gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc
     4501 tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg
     4561 ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc
     4621 cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ctgagtcgac cgcccgcccc
     4681 acgacccgca gcgcccgacc gaaaggagcg cacgacccca tggctcgacg ataaaataaa
     4741 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     4801 agctagctta agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaga
     4861 agttcagatc aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg
     4921 tggtaagcag ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc
     4981 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc
     5041 cccagatgcg gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc
     5101 ccaaggacct gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct
     5161 tctgttcgcg cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg
     5221 gcgccagtcc tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg
     5281 cagttgcatc cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac
     5341 tacccgtcag cgggggtctt tcacatgcag catgtatcaa aattaatttg gttttttttc
     5401 ttaagtattt acattaaatg gccatagttg cattaatgaa tcggccaacg cgcggggaga
     5461 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     5521 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     5581 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     5641 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5701 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5761 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5821 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5881 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5941 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6001 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6061 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6121 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6181 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6241 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6301 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6361 ttgcggccgg ccgcaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6421 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6481 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6541 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     6601 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     6661 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     6721 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     6781 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     6841 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     6901 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     6961 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7021 gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7081 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7141 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7201 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7261 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7321 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7381 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     7441 cctataaaaa taggcgtatc acgaggccct ttcgtcttca ctataggagc ttggcccatt
     7501 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
     7561 gccatgtt
//
