LOCUS       sequence-80499-e        6328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a bicistronic IRES-mAtetrine containing retroviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6328)
  TITLE     Vignali lab plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6328)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pMSCV-IRES-mAmetrine 1.1 FP
FEATURES             Location/Qualifiers
     source          1..6328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1473..2022
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1617..1634)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1844..1863
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(2025..2027,2028..2030,2031..2744)
                     /codon_start=1
                     /product="photostable yellow fluorescent protein with a 
                     large Stokes shift, excitable with violet light (Ai et al.,
                     2008)"
                     /label=mAmetrine1.1
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVRGEGEGDATNGKLTL
                     KFICTSGELPVPWPTLVTTLSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GSYRTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNMNAWDAYITADKQKNGIK
                     ANFKIEHNVKDGGVQLADAYQQNTPIGDGSVLLPDNHYLSFQSKLFKDPNEQRDHMVLL
                     EFVTAAGITPGMDELYK"
     primer_bind     complement(2070..2091)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3436..3452)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3436..3452)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3449..3471)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3460..3476
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3484..3490,3491..3508,3509..3514))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3529..3550
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3667..3684)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3838..4426)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3918..3937)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4597..5388,5389..5457))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5220..5239
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5458..5562)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5630..5648
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5686..5708)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5808..5827
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6021..6043
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6035..6052
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6036..6052
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6251..6270
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
      121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
      181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
      241 ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt cagatcttac gtagctagcg gatcccaatt
     1441 gctcgagcgg gatcaattcc gccccccccc taacgttact ggccgaagcc gcttggaata
     1501 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt
     1561 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct
     1621 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc
     1681 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga
     1741 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc
     1801 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt
     1861 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg
     1921 gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc
     1981 gaaccacggg gacgtggttt tcctttgaaa aacacgataa taccatggtg agcaagggcg
     2041 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2101 acaggttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag ctgaccctga
     2161 agttcatctg cacctccggc gagctgcccg tgccctggcc caccctcgtg accaccctgt
     2221 cttacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca
     2281 agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctccttcaag gacgacggca
     2341 gctacaggac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2401 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaaca
     2461 tgaacgcgtg ggacgcgtat atcacggccg acaagcagaa gaacggcatc aaagcgaact
     2521 tcaagatcga gcacaacgtc aaggacggcg gcgtgcagct cgccgacgcg taccagcaga
     2581 acacccccat cggcgacggc tccgtgctgc tgcctgacaa ccactacctg agcttccaga
     2641 gcaagctgtt caaagacccc aacgagcagc gcgatcacat ggtcctgctg gagttcgtta
     2701 ccgccgccgg gataactccc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
     2761 gagtcgacct gcaggcatgc aagcttcagg tagccggcta acgttaacaa ccggtacctc
     2821 tagaactata gctagcatgc gcaaatttaa agcgctgata tcgataaaat aaaagatttt
     2881 atttagtctc cagaaaaagg ggggaatgaa agaccccacc tgtaggtttg gcaagctagc
     2941 ttaagtaacg ccattttgca aggcatggaa aatacataac tgagaataga gaagttcaga
     3001 tcaaggttag gaacagagag acagcagaat atgggccaaa caggatatct gtggtaagca
     3061 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc cgccctcagc
     3121 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa atgaccctgt
     3181 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     3241 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     3301 gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct
     3361 cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca
     3421 gaattggtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3481 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3541 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3601 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3661 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3721 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3781 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3841 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3901 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3961 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4021 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4081 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4141 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4201 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4261 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4321 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4381 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4441 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4501 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4561 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4621 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4681 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4741 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4801 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     4861 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     4921 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     4981 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5041 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5101 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5161 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5221 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5281 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5341 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5401 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5461 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5521 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5581 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     5641 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     5701 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     5761 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     5821 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     5881 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5941 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     6001 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccacgct
     6061 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     6121 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     6181 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     6241 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     6301 tgccggccac gatgcgtccg gcgtagag
//
