LOCUS       sequence-328859-        4800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To target Venus to the mitochondrial outer membrane. MAO tail anchor
            inserts into the outer membrane and Venus faces the cytosol. Serves 
            as a collisional control in FLIM-FRET experiments..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4800)
  TITLE     Andrew lab Venus and Cerulean fusions
REFERENCE   2  (bases 1 to 4800)
  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-7aa-MAO-pEGFP-C3
FEATURES             Location/Qualifiers
     source          1..4800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..313
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        314..517
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     468..488
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      556..565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(562..564,565..567,568..1278)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     ERVTAAGITLGMDELYK"
     primer_bind     complement(607..628)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(868..887)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1537..1658
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1574..1593)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1628..1647
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1665..2120)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1802..1823)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2014..2033
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2147..2251
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2253..2610
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2253..2444
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2276..2296)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2461..2596
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2645..3439
                     /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             2645..3439
                     /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(2699..2718)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3309..3328
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3627..3646)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3671..3718
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4047..4635
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4536..4555
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
       61 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
      121 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
      181 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
      241 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
      301 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
      361 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
      421 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
      481 taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc gtcagatccg
      541 ctagcgctac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      601 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      661 ggcgagggcg atgccaccta cggcaagctg accctgaagc tgatctgcac caccggcaag
      721 ctgcccgtgc cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc
      781 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      841 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      901 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      961 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1021 accgccgaca agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag
     1081 gacggcggcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1141 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     1201 gagaagcgcg atcacatggt cctgctggaa cgcgtgaccg ccgccgggat cactctcggc
     1261 atggacgagc tgtacaagtc cggactcaga tctcgagttt gggaaaggaa cctgccctct
     1321 gtttctggcc tgctgaagat cattggattt tccacatcag taactgccct ggggtttgtg
     1381 ctgtacaaat acaagctcct gccacggtct taagtcgact ctagataact gatcataatc
     1441 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     1501 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     1561 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     1621 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     1681 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     1741 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     1801 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     1861 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     1921 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     1981 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     2041 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     2101 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2161 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2221 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     2281 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2341 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     2401 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     2461 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     2521 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     2581 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     2641 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     2701 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     2761 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     2821 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     2881 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     2941 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3001 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3061 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3121 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     3181 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3241 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     3301 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     3361 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     3421 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     3481 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     3541 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     3601 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3661 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     3721 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     3781 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     3841 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     3901 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     3961 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4021 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4081 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4141 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4201 gtgtagccgt agttaggcca ccacttcaag aactctgtat caccgcctac atacctcgct
     4261 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4321 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4381 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4441 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4501 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4561 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4621 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     4681 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     4741 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
//
