LOCUS       sequence-364104-        7566 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Stable expression of HaloTag7-Galectin3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7566)
  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 7566)
  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-HaloTag7-Galectin3
FEATURES             Location/Qualifiers
     source          1..7566
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    747..1104
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1169..1585
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1515..1537
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1553..1569
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1595..1969
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1907..1923
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     regulatory      1996..2005
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2002..2889
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     misc_feature    3710..4276
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3726..4278
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3870..3887)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4097..4116
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4296..4670
                     /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             4900..5369
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5499..5516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6422..7213,7214..7282))
                     /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     7045..7064
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7283..7387)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7455..7473
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatgct agagaaccat cagatgtttc cagggtgccc
      421 caaggacctg aaatgaccct gtgccttatt tgaactaacc aatcagttcg cttctcgctt
      481 ctgttcgcgc gcttctgctc cccgagctca ataaaagagc ccacaacccc tcactcggcg
      541 cgccagtcct ccgattgact gagtcgcccg ggtacccgtg tatccaataa accctcttgc
      601 agttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga gtgattgact
      661 acccgtcagc gggggtcttt catttggggg ctcgtccggg atcgggagac ccctgcccag
      721 ggaccaccga cccaccaccg ggaggtaagc tggccagcaa cttatctgtg tctgtccgat
      781 tgtctagtgt ctatgactga ttttatgcgc ctgcgtcggt actagttagc taactagctc
      841 tgtatctggc ggacccgtgg tggaactgac gagttcggaa cacccggccg caaccctggg
      901 agacgtccca gggacttcgg gggccgtttt tgtggcccga cctgagtcca aaaatcccga
      961 tcgttttgga ctctttggtg cacccccctt agaggaggga tatgtggttc tggtaggaga
     1021 cgagaaccta aaacagttcc cgcctccgtc tgaatttttg ctttcggttt gggaccgaag
     1081 ccgcgccgcg cgtcttgtct gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt
     1141 tctgtatttg tctgaaaata tgggcccggg ccagactgtt accactccct taagtttgac
     1201 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
     1261 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
     1321 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
     1381 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
     1441 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
     1501 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
     1561 agccctcact ccttctctag gcgcccccat atggccatat gagatcttat atggggcacc
     1621 cccgcccctt gtaaacttcc ctgaccctga catgacaaga gttactaaca gcccctctct
     1681 ccaagctcac ttacaggctc tctacttagt ccagcacgaa gtctggagac ctctggcggc
     1741 agcctaccaa gaacaactgg accgaccggt ggtacctcac ccttaccgag tcggcgacac
     1801 agtgtgggtc cgccgacacc agactaagaa cctagaacct cgctggaaag gaccttacac
     1861 agtcctgctg accaccccca ccgccctcaa agtagacggc atcgcagctt ggatacacgc
     1921 cgcccacgtg aaggctgccg accccggggg tggaccatcc tctagactgc cggatctgcc
     1981 atcgatataa agcttgccac catggaaatc ggtactggct ttccattcga cccccattat
     2041 gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga tggcacccct
     2101 gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat catcccgcat
     2161 gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa atccgacaaa
     2221 ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt catcgaagcc
     2281 ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct gggtttccac
     2341 tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt catccgccct
     2401 atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc cttccgcacc
     2461 accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg tacgctgccg
     2521 atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga gccgttcctg
     2581 aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat cgccggtgag
     2641 ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca gtcccctgtc
     2701 ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga agccgctcgc
     2761 ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccaggtct gaatctgctg
     2821 caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc tactctggag
     2881 atttccggtg gttcaggctc gggatccggg aattcgggta tggcagacaa tttttcgctc
     2941 catgatgcgt tatctgggtc tggaaaccca aaccctcaag gatggcctgg cgcatggggg
     3001 aaccagcctg ctggggcagg gggctaccca ggggcttcct atcctggggc ctaccccggg
     3061 caggcacccc caggggctta tcctggacag gcacctccag gcgcctaccc tggagcacct
     3121 ggagcttatc ccggagcacc tgcacctgga gtctacccag ggccacccag cggccctggg
     3181 gcctacccat cttctggaca gccaagtgcc cccggagcct accctgccac tggcccctat
     3241 ggcgcccctg ctgggccact gattgtgcct tataacctgc ctttgcctgg gggagtggtg
     3301 cctcgcatgc tgataacaat tctgggcacg gtgaagccca atgcaaacag aattgcttta
     3361 gatttccaaa gagggaatga tgttgccttc cactttaacc cacgcttcaa tgagaacaac
     3421 aggagagtca ttgtttgcaa tacaaagctg gataataact ggggaaggga agaaagacag
     3481 tcggttttcc catttgaaag tgggaaacca ttcaaaatac aagtactggt tgaacctgac
     3541 cacttcaagg ttgcagtgaa tgatgctcac ttgttgcagt acaatcatcg ggttaaaaaa
     3601 ctcaatgaaa tcagcaaact gggaatttct ggtgacatag acctcaccag tgcttcatat
     3661 accatgatat aagaattcgg gtctcgagta aagcggccgc tacgtaaatt ccgccccccc
     3721 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     3781 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     3841 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     3901 tgtcgtgaag gaagcagttc ctctggaagt ttcttgaaga caaacaacgt ctgtagcgac
     3961 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     4021 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     4081 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     4141 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     4201 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     4261 aaaaacacga tgataatatg gccacaaccg tcgacatggc caagttgacc agtgccgttc
     4321 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     4381 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     4441 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     4501 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     4561 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     4621 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga gtcgaccgcc
     4681 cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatggc tcgacgataa
     4741 aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt
     4801 ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca taactgagaa
     4861 tagagaagtt cagatcaagg tcaggaacag atggaacagc tgaatatggg ccaaacagga
     4921 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatgga acagctgaat
     4981 atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag
     5041 atggtcccca gatgcggtcc agccctcagc agtttctaga gaaccatcag atgtttccag
     5101 ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga actaaccaat cagttcgctt
     5161 ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata aaagagccca caacccctca
     5221 ctcggggcgc cagtcctccg attgactgag tcgcccgggt acccgtgtat ccaataaacc
     5281 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
     5341 attgactacc cgtcagcggg ggtctttcac atgcagcatg tatcaaaatt aatttggttt
     5401 tttttcttaa gtatttacat taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg
     5461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6361 atccttttgc ggccggccgc aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6421 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6481 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6541 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6601 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6661 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6721 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6781 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6841 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6901 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6961 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7021 gctcttgccc ggcgtcaaca cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7081 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7141 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7201 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7261 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7321 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7381 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7441 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcactat aggagcttgg
     7501 cccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
     7561 attacc
//
