LOCUS       sequence-347649-        4914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To increase the length of the linker between Venus and Noxa-4E 
            proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4914)
  TITLE     Andrew lab Venus and Cerulean fusions
REFERENCE   2  (bases 1 to 4914)
  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"
COMMENT     Sequence Label: Venus-30aa-Noxa-4E-pEGFP-C1
FEATURES             Location/Qualifiers
     source          1..4914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        87..390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        391..594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     545..565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      633..642
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(639..641,642..644,645..1355)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     VNFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     ERVTAAGITHGMDELYK"
     primer_bind     complement(684..705)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(945..964)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1728..1849
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1765..1784)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1819..1838
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1856..2311)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1993..2014)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2205..2224
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2338..2442
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2444..2801
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2444..2635
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2467..2487)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2652..2787
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2836..3630
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2836..3630
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2890..2909)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3500..3519
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3818..3837)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3862..3909
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4238..4826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4727..4746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      421 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      481 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      541 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      601 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      661 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      721 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagctga
      781 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgggctacg
      841 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      901 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      961 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1021 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1081 gccacaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggtc aacttcaaga
     1141 tccgccacaa catcgaggac ggcggcgtgc agctcgccga ccactaccag cagaacaccc
     1201 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagctac cagtccaagc
     1261 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggaacgc gtgaccgccg
     1321 ccgggatcac tcacggcatg gacgagctgt acaagtccgg actcactagt ggcgccctcg
     1381 agacgcgtcc cgggggtacc gctggaccgg tcgccggcga gacgcgtggc agatctcgag
     1441 gaggcatgcc tgggaagaag gcgcgcaaga acgctcaacc gagccccgcg cgggctccag
     1501 cagagctgga agtcgaggaa gctactcaag agaggagaga aggagacaaa gagaacttcc
     1561 ggcagaaact tctgaatctg atatccaaac tcttctgctc aggaacctga tctagataac
     1621 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     1681 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     1741 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     1801 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     1861 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     1921 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     1981 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     2041 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     2101 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     2161 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     2221 ggagcgggcg ctagggcgct ggcaattgta gcggtcacgc tgcgcgtaac caccacaccc
     2281 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     2341 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2401 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     2461 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2521 gtatacaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     2581 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     2641 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     2701 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     2761 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     2821 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2881 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2941 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3001 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3061 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3121 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3181 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3241 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3301 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3361 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3421 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3481 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3541 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3601 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3661 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3721 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3781 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     3841 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     3901 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     3961 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     4021 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     4081 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4141 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4201 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4261 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     4321 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     4381 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta tcaccgccta
     4441 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4501 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4561 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4621 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4681 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4741 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4801 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4861 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctg
//
