LOCUS       sequence-501102-        7688 bp    DNA     circular SYN 25-FEB-2026
DEFINITION  Retroviral expression of ALFA-mouse Fis1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7688)
  TITLE     David Chan Lab Plasmids Jan 2026
REFERENCE   2  (bases 1 to 7688)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 25, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pQCXIP ALFA-mouse Fis1
FEATURES             Location/Qualifiers
     source          1..7688
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        309..512
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             513..688
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    751..950
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1151..1567
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1497..1519
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1535..1551
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1604..1907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1908..2111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2108..2132
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2265..2303
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     misc_feature    2821..3388
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2982..2999)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3209..3228
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3421..4020
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3421..3440)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3917..3937
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4394..4819
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(4922..4944)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5080..5100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5085..5414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5265..5400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5436..5455
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5536..5553)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5707..6295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5787..5806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6466..7257,7258..7326))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7089..7108
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7327..7431)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7499..7517
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(7617..7688,1..308)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
       61 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      301 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      361 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      421 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      481 gtgtacggtg ggaggtctat ataagcagag ctcaataaaa gagcccacaa cccctcactc
      541 ggcgcgccag tcttccgata gactgcgtcg cccgggtacc cgtattccca ataaagcctc
      601 ttgctgtttg catccgaatc gtggtctcgc tgttccttgg gagggtctcc tctgagtgat
      661 tgactaccca cgacgggggt ctttcatttg ggggctcgtc cgggatttgg agacccctgc
      721 ccagggacca ccgacccacc accgggaggt aagctggcca gcaacttatc tgtgtctgtc
      781 cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt ctgtactagt tagctaacta
      841 gctctgtatc tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc
      901 tgggagacgt cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc
      961 gatgtggaat ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca
     1021 gttcccgcct ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct
     1081 gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc
     1141 tgaaaattag ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga
     1201 tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt
     1261 ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg
     1321 agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga
     1381 ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa
     1441 gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct
     1501 tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct
     1561 aggcgccgga attgaagatc tgggggatcg atcctctaga gtccgttaca taacttacgg
     1621 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     1681 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     1741 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     1801 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1861 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1921 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1981 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     2041 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     2101 taagcagagc tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg
     2161 acctccatag aagacaccgg gaccgatcca gcctccgcgg ccgggaacgg tgcattggaa
     2221 cggacctgca ggaattgatc cgcggccgct aagccaccat gccctcccgg ctggaggagg
     2281 agctgcggcg gcggctgacc gagtccggcc ggactcagat ctcgagctca agcttcgaat
     2341 tcatggaggc cgtgctgaac gagctggtgt ctgtggagga tctgaagaat tttgaaagga
     2401 aatttcagtc tgagcaggca gctggttctg tgtccaagag cacgcaattt gaatatgcct
     2461 ggtgcctggt tcgaagcaaa tacaatgagg acatccgcag aggcatcgtg ctgctggagg
     2521 agctgttgcc caaagggagc aaagaggaac agcgggacta tgtcttctac ctggccgtgg
     2581 gcaactaccg gctcaaggaa tatgaaaagg ctctaaagta tgtgcgaggg ctgttgcaga
     2641 ctgagcccca gaacaaccag gccaaggagc tggaacgcct gattgataag gccatgaaga
     2701 aagatggact ggtaggcatg gccatcgttg gtggcatggc cctgggcgtg gcaggcctgg
     2761 ctggactcat tggactggct gtctccaagt ccaaatcctg aggatccaag ctgtaggaat
     2821 tccgcccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt
     2881 tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc
     2941 tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca
     3001 aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac
     3061 gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg
     3121 ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt
     3181 gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct
     3241 gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg
     3301 ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt
     3361 ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccacaaggag acgaccttcc
     3421 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta
     3481 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac
     3541 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     3601 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     3661 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     3721 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     3781 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     3841 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     3901 gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     3961 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     4021 cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca tggctccgac
     4081 cgaagccacc cggggcggcc ccgccgaccc cgcacccgcc cccgaggccc accgactcta
     4141 gtcgagggct gcagcgctgc agaggccgag tgcagaactg ctccaaaggg acctcaaggc
     4201 tttccgaggg acactaggct gactccatcg agccagtgta gagataagct tatcgattag
     4261 tccaatttgt taaagacagg atatcagtgg tccaggctct agttttgact caacaatatc
     4321 accagctgaa gcctatagag tacgagccat agataaaata aaagatttta tttagtctcc
     4381 agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct taagtaacgc
     4441 cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga tcaaggtcag
     4501 gaacagatgg aacagggtcg accctagaga accatcagat gtttccaggg tgccccaagg
     4561 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4621 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     4681 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4741 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4801 tcagcggggg tctttcattt gggggctcgt ccgggatcgg gagacccctg cccagggacc
     4861 accgacccac caccgggagg taagctggct gcctcgcgcg tttcggtgat gacggtgaaa
     4921 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     4981 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     5041 cccagtcacg tagcgatagc ggagtgtaga tccggctgtg gaatgtgtgt cagttagggt
     5101 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     5161 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     5221 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     5281 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     5341 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     5401 taggcttttg caaaaagctt actggcttaa ctatgcggca tcagagcaga ttgtactgag
     5461 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     5521 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5581 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5641 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6121 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     6481 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     6541 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     6601 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     6661 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     6721 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6781 tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6841 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6901 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6961 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7021 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7081 aacacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7141 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7201 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7261 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7321 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     7381 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     7441 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     7501 taggcgtatc acgaggccct ttcgtcttca agaattagct tggccattgc atacgttgta
     7561 tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc catgttgaca
     7621 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     7681 tatggagt
//
