LOCUS       sequence-379670-        6465 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of YFP-Bak chimera.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6465)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6465)
  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            "unpublished"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-YFP-Bak
FEATURES             Location/Qualifiers
     source          1..6465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(229..251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     351..370
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     564..586
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     714..733
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             933..1449
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    1513..1854
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             1921..2337
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     2267..2289
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     2305..2321
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2362..2371
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2368..2370,2371..2373,2374..3084)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2413..2434)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2674..2693)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3021..3042
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3792..4306
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4468..4484
                     /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(4492..4498,4499..4516,4517..4522))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4537..4558
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4675..4692)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4846..5434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4926..4945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5605..6396,6397..6465))
                     /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     6228..6247
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      361 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      421 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
      481 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      541 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat
      601 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
      661 acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg
      721 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
      781 ccggcgtaga ggcgattagt ccaatttgtt aaagacagga tatcagtggt ccaggctcta
      841 gttttgactc aacaatatca ccagctgaag cctatagagt acgagccata gataaaataa
      901 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
      961 aagctagctt aagtaacgcc attttgcaag gcatggaaaa tacataactg agaatagaga
     1021 agttcagatc aaggttagga acagagagac agcagaatat gggccaaaca ggatatctgt
     1081 ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtcccg
     1141 ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg
     1201 accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc
     1261 tgctccccga gctcaataaa agagcccaca acccctcact cggcgcgcca gtcctccgat
     1321 agactgcgtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     1381 cgtggactcg ctgatccttg ggagggtctc ctcagattga ttgactgccc acctcggggg
     1441 tctttcattt ggaggttcca ccgagatttg gagacccctg cccagggacc accgaccccc
     1501 ccgccgggag gtaagctggc cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt
     1561 tgtgccggca tctaatgttt gcgcctgcgt ctgtactagt tagctaacta gctctgtatc
     1621 tggcggaccc gtggtggaac tgacgagttc tgaacacccg gccgcaaccc tgggagacgt
     1681 cccagggact ttgggggccg tttttgtggc ccgacctgag gaagggagtc gatgtggaat
     1741 ccgaccccgt caggatatgt ggttctggta ggagacgaga acctaaaaca gttcccgcct
     1801 ccgtctgaat ttttgctttc ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc
     1861 tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc tgtatttgtc tgaaaattag
     1921 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
     1981 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
     2041 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
     2101 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
     2161 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
     2221 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
     2281 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccgga
     2341 attcgttaac ctcgaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     2401 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     2461 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     2521 ggcaagctgc ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgcagtgc
     2581 ttcgcccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     2641 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     2701 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     2761 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     2821 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     2881 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     2941 ggccccgtgc tgctgcccga caaccactac ctgagctacc agtccgccct gagcaaagac
     3001 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     3061 ctcggcatgg acgagctgta caagtccgga ctcagatctg catctggaca aggaccaggt
     3121 cccccgaagg tgggctgcga tgagtccccg tccccttctg aacagcaggt tgcccaggac
     3181 acagaggagg tctttcgaag ctacgttttt tacctccacc agcaggaaca ggagacccag
     3241 ggggcggccg cccctgccaa ccccgagatg gacaacttgc ccctggaacc caacagcatc
     3301 ttgggtcagg tgggtcggca gcttgctctc atcggagatg atattaaccg gcgctacgac
     3361 acagagttcc agaatttact agaacagctt cagcccacag ccgggaatgc ctacgaactc
     3421 ttcaccaaga tcgcctccag cctatttaag agtggcatca gctggggccg cgtggtggct
     3481 ctcctgggct ttggctaccg tctggccctg tacgtctacc agcgtggttt gaccggcttc
     3541 ctgggccagg tgacctgctt tttggctgat atcatactgc atcattacat cgccagatgg
     3601 atcgcacaga gaggcggttg ggtggcagcc ctgaatttgc gtagagaccc catcctgacc
     3661 gtaatggtga tttttggtgt ggttctgttg ggccaattcg tggtacacag attcttcaga
     3721 tcatgaagat ctcgagctca agcttatcga taaaataaaa gattttattt agtctccaga
     3781 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     3841 tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac
     3901 agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     3961 tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac
     4021 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     4081 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     4141 agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc
     4201 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     4261 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct
     4321 tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat
     4381 tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg
     4441 cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac acaacatacg
     4501 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     4561 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     4621 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4681 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4741 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4801 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4861 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4921 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4981 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5041 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5101 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5161 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5221 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5281 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5341 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5401 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5461 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5521 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5581 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5641 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5701 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5761 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5821 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5881 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5941 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     6001 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     6061 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     6121 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     6181 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     6241 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     6301 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     6361 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6421 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcat
//
