LOCUS       sequence-501133-        8382 bp    DNA     circular SYN 25-FEB-2026
DEFINITION  Retroviral expression of EGFP-mouse Fis1 (L110A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8382)
  TITLE     David Chan Lab Plasmids Jan 2026
REFERENCE   2  (bases 1 to 8382)
  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 EGFP-mouse Fis1 (L110A)
FEATURES             Location/Qualifiers
     source          1..8382
                     /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"
     regulatory      2250..2259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2256..2258,2259..2261,2262..2972)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2256..2258,2259..2261,2262..2972)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2301..2322)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2562..2581)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2909..2930
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    join(3509..4083,4084..4086,4087..4095)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3676..3693)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3903..3922
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4115..4714
                     /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(4115..4134)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4611..4631
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5088..5513
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     primer_bind     complement(5616..5638)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5774..5794)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5779..6108
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5959..6094
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6130..6149
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6230..6247)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6401..6989)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6481..6500)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7160..7951,7952..8020))
                     /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     7783..7802
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8021..8125)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8193..8211
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        join(8311..8382,1..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 cgcggccgcg ccaccatggt gagcaagggc gaggagctgt
     2281 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     2341 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
     2401 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     2461 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     2521 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
     2581 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     2641 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
     2701 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     2761 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     2821 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     2881 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     2941 ggatcactct cggcatggac gagctgtaca agtccggccg gactcagatc tcgagctcaa
     3001 gcttcgaatt catggaggcc gtgctgaacg agctggtgtc tgtggaggat ctgaagaatt
     3061 ttgaaaggaa atttcagtct gagcaggcag ctggttctgt gtccaagagc acgcaatttg
     3121 aatatgcctg gtgcctggtt cgaagcaaat acaatgagga catccgcaga ggcatcgtgc
     3181 tgctggagga gctgttgccc aaagggagca aagaggaaca gcgggactat gtcttctacc
     3241 tggccgtggg caactaccgg ctcaaggaat atgaaaaggc tctaaagtat gtgcgagggc
     3301 tgttgcagac tgagccccag aacaaccagg ccaaggaggc agaacgcctg attgataagg
     3361 ccatgaagaa agatggactg gtaggcatgg ccatcgttgg tggcatggcc ctgggcgtgg
     3421 caggcctggc tggactcatt ggactggctg tctccaagtc caaatcctga ggatccaagc
     3481 tgtaggaatt ccsccccccc cccccccccc cctctccctc ccccccccct aacgttactg
     3541 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat
     3601 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc
     3661 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag
     3721 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc
     3781 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac
     3841 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca
     3901 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt
     3961 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa
     4021 aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa acacgatgat
     4081 aagcttgcca caacccacaa ggagacgacc ttccatgacc gagtacaagc ccacggtgcg
     4141 cctcgccacc cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga
     4201 ctaccccgcc acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct
     4261 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     4321 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4381 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4441 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4501 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4561 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4621 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4681 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     4741 gaccgaaagg agcgcacgac cccatggctc cgaccgaagc cacccggggc ggccccgccg
     4801 accccgcacc cgcccccgag gcccaccgac tctagtcgag ggctgcagcg ctgcagaggc
     4861 cgagtgcaga actgctccaa agggacctca aggctttccg agggacacta ggctgactcc
     4921 atcgagccag tgtagagata agcttatcga ttagtccaat ttgttaaaga caggatatca
     4981 gtggtccagg ctctagtttt gactcaacaa tatcaccagc tgaagcctat agagtacgag
     5041 ccatagataa aataaaagat tttatttagt ctccagaaaa aggggggaat gaaagacccc
     5101 acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg gaaaaataca
     5161 taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagg gtcgacccta
     5221 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     5281 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     5341 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     5401 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     5461 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc atttgggggc
     5521 tcgtccggga tcgggagacc cctgcccagg gaccaccgac ccaccaccgg gaggtaagct
     5581 ggctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     5641 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     5701 gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg
     5761 tagatccggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     5821 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     5881 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     5941 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     6001 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
     6061 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttactggc
     6121 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     6181 cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg
     6241 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     6301 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     6361 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     6421 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     6481 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     6541 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     6601 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     6661 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6721 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     6781 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6841 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6901 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6961 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     7021 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     7081 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     7141 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     7201 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     7261 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     7321 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     7381 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     7441 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
     7501 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     7561 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     7621 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     7681 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     7741 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
     7801 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     7861 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     7921 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     7981 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     8041 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     8101 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag
     8161 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
     8221 ttcaagaatt agcttggcca ttgcatacgt tgtatccata tcataatatg tacatttata
     8281 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt
     8341 aatcaattac ggggtcatta gttcatagcc catatatgga gt
//
