LOCUS       Exported                5450 bp ds-DNA     circular SYN 27-APR-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5450)
  AUTHORS   Koushik SV, Chen H, Thaler C, Puhl HL, Vogel SS
  TITLE     Cerulean, Venus, and VenusY67C FRET reference standards.
  JOURNAL   Biophys J. 2006 Dec 15. 91(12):L99-L101.
  PUBMED    17040988
REFERENCE   2  (bases 1 to 5450)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 29, 2022 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5450
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          532..548
                     /organism="pLenti CMV GFP Puro (658-5)"
                     /mol_type="other DNA"
     source          549..1295
                     /organism="pLenti CMV GFP Puro (658-5)"
                     /mol_type="other DNA"
     source          1296..1315
                     /organism="pLenti CMV GFP Puro (658-5)"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     506..551
                     /label=A-P1
     misc_feature    543..551
                     /label=Kozak
     CDS             549..1295
                     /codon_start=1
                     /label=NDUFV2
                     /translation="MFFSAALRARAAGLTAHWGRHVRNLHKTVMQNGAGGALFVHRDTP
                     ENNPDTPFDFTPENYKRIEAIVKNYPEGHKAAAVLPVLDLAQRQNGWLPISAMNKVAEV
                     LQVPPMRVYEVATFYTMYNRKPVGKYHIQVCTTTPCMLRNSDSILEAIQKKLGIKVGET
                     TPDKLFTLIEVECLGACVNAPMVQINDNYYEDLTAKDIEEIIDELKAGKIPKPGPRSGR
                     FSCEPAGGLTSLTEPPKGPGFGVQAGL"
     primer_bind     complement(1296..1340)
                     /label=B-P2
     CDS             1296..1298
                     /codon_start=1
                     /label=mutated stop
                     /translation="L"
     CDS             1299..1337
                     /codon_start=1
                     /label=13aa linker
                     /translation="NIGGISNSAVDGT"
     CDS             1338..2048
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKL
                     ICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     polyA_signal    2178..2299
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2215..2234)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2269..2288
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2306..2761)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2443..2464)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2655..2674
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2788..2892
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2894..3251
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2917..2937)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3102..3237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3164..3183
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3187..3198)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3286..4080
                     /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(3340..3359)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3950..3969
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4268..4287)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4312..4359
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4688..5276
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5177..5196
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gccgactcta
      541 gagccaccat gttcttctcc gcggcgctcc gggcccgggc ggctggcctc accgcccact
      601 ggggaagaca tgtaaggaat ttgcataaga cagttatgca aaatggagct ggaggagctt
      661 tatttgtgca cagagatact cctgagaata accctgatac tccatttgat ttcacaccag
      721 aaaactataa gaggatagag gcaattgtaa aaaactatcc agaaggccat aaagcagcag
      781 ctgttcttcc agtcctggat ttagcccaaa ggcagaatgg gtggttgccc atctctgcta
      841 tgaacaaggt tgcagaagtt ttacaagtac ctccaatgag agtatatgaa gtagcaactt
      901 tttatacaat gtataatcga aagccagttg gaaagtatca cattcaggtc tgcactacta
      961 caccctgcat gcttcgaaac tctgacagca tactggaggc cattcagaaa aagcttggaa
     1021 taaaggttgg ggagactaca cctgacaaac ttttcactct tatagaagtg gaatgtttag
     1081 gggcctgtgt gaacgcacca atggttcaaa taaatgacaa ttactatgag gatttgacag
     1141 ctaaggatat tgaagaaatt attgatgagc tcaaggctgg caaaatccca aaaccagggc
     1201 caaggagtgg acgcttctct tgtgagccag ctggaggtct tacctctttg actgaaccac
     1261 ccaagggacc tggatttggt gtacaagcag gccttttaaa cattggtggg atatcaaatt
     1321 ctgcagtcga cggtaccagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     1381 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     1441 atgccaccta cggcaagctg accctgaagc tcatctgcac caccggcaag ctgcccgtgc
     1501 cctggcccac cctcgtgacc accctcggct acggcctgca gtgcttcgcc cgctaccccg
     1561 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1621 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1681 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1741 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca
     1801 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg
     1861 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1921 ccgacaacca ctacctgagc taccagtcca agctgagcaa agaccccaac gagaagcgcg
     1981 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     2041 tgtacaaggg atccaccgga tctagataac tgatcataat cagccatacc acatttgtag
     2101 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2161 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2221 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2281 aactcatcaa tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     2341 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     2401 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     2461 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     2521 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     2581 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2641 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2701 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2761 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2821 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2881 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2941 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3001 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3061 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     3121 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     3181 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     3241 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3301 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3361 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3421 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     3481 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3541 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3601 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3661 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3721 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3781 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3841 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3901 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3961 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4021 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4081 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4141 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4201 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     4261 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     4321 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     4381 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     4441 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     4501 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     4561 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4621 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4681 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4741 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4801 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4861 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4921 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4981 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5041 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5101 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5161 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5221 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5281 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5341 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     5401 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
