LOCUS       Exported                5087 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 5087)
  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 5087)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 29, 2022 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5087
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          532..548
                     /organism="pLenti CMV GFP Puro (658-5)"
                     /mol_type="other DNA"
     source          549..932
                     /organism="pLenti CMV GFP Puro (658-5)"
                     /mol_type="other DNA"
     source          933..952
                     /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..932
                     /codon_start=1
                     /label=NDUFB6
                     /translation="MTGYTPDEKLRLQQLRELRRRWLKDQELSPREPVLPPQKMGPMEK
                     FWNKFLENKSPWRKMVHGVYKKSIFVFTHVLVPVWIIHYYMKYHVSEKPYGIVEKKSRI
                     FPGDTILETGEVIPPMKEFPDQHH"
     primer_bind     complement(933..977)
                     /label=B-P2
     CDS             933..935
                     /codon_start=1
                     /label=mutated stop
                     /translation="L"
     CDS             936..974
                     /codon_start=1
                     /label=13aa linker
                     /translation="NIGGISNSAVDGT"
     CDS             975..1685
                     /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    1815..1936
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1852..1871)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1906..1925
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1943..2398)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2080..2101)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2292..2311
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2425..2529
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2531..2888
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2554..2574)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2739..2874
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2801..2820
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2824..2835)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2923..3717
                     /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(2977..2996)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3587..3606
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3905..3924)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3949..3996
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4325..4913
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4814..4833
                     /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 gacggggtac actccggatg agaaactgcg gctgcagcag ctgcgagagc
      601 tgagaaggcg atggctgaag gaccaggagc tgagccctcg ggagccggtg ctgcccccac
      661 agaagatggg gcctatggag aaattctgga ataaattttt ggagaataaa tccccttgga
      721 ggaaaatggt ccatggggta tacaaaaaga gtatctttgt tttcactcat gtacttgtac
      781 ctgtctggat tattcattat tacatgaagt atcatgtttc tgaaaaacca tatggcatag
      841 ttgaaaagaa gtccagaata ttccctggtg atacaattct ggagactgga gaagtaattc
      901 caccaatgaa agaatttcct gatcaacatc atttaaacat tggtgggata tcaaattctg
      961 cagtcgacgg taccagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1021 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1081 ccacctacgg caagctgacc ctgaagctca tctgcaccac cggcaagctg cccgtgccct
     1141 ggcccaccct cgtgaccacc ctcggctacg gcctgcagtg cttcgcccgc taccccgacc
     1201 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1261 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     1321 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     1381 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcacc gccgacaagc
     1441 agaagaacgg catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcggcgtgc
     1501 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1561 acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc
     1621 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     1681 acaagggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     1741 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     1801 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1861 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1921 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     1981 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     2041 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     2101 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     2161 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     2221 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     2281 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     2341 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     2401 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2461 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2521 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2581 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2641 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2701 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2761 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2821 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2881 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2941 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     3001 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     3061 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     3121 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     3181 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     3241 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3301 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3361 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3421 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     3481 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3541 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3601 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3661 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3721 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3781 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3841 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     3901 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     3961 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     4021 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     4081 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     4141 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     4201 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     4261 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     4321 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     4381 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     4441 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     4501 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     4561 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     4621 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     4681 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     4741 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     4801 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4861 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4921 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4981 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttattaatag
     5041 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccg
//
