LOCUS       Exported                5051 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 5051)
  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 5051)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Apr 29, 2022 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5051
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          532..551
                     /organism="pLenti CMV GFP Puro (658-5)"
                     /mol_type="other DNA"
     source          552..896
                     /organism="pLenti CMV GFP Puro (658-5)"
                     /mol_type="other DNA"
     source          897..916
                     /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..896
                     /codon_start=1
                     /label=NDUFA5
                     /translation="MAGVLKKTTGLVGLAVCNTPHERLRILYTKILDVLEEIPKNAAYR
                     KYTEQITNEKLAMVKAEPDVKKLEDQLQGGQLEEVILQAEHELNLARKMREWKLWEPLV
                     EEPPADQWKWPI"
     primer_bind     complement(897..941)
                     /label=B-P2
     CDS             897..899
                     /label=mutated stop
     CDS             900..938
                     /codon_start=1
                     /label=13aa linker
                     /translation="NIGGISNSAVDGT"
     CDS             939..1649
                     /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    1779..1900
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1816..1835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1870..1889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1907..2362)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2044..2065)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2256..2275
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2389..2493
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2495..2852
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2518..2538)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2703..2838
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2765..2784
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2788..2799)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2887..3681
                     /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(2941..2960)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3551..3570
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3869..3888)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3913..3960
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4289..4877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4778..4797
                     /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 ggcgggtgtg ctgaagaaga ccactggcct tgtgggattg gctgtgtgca
      601 atactcctca cgagaggcta agaatattgt acacaaagat tcttgatgtt cttgaggaaa
      661 tccctaaaaa tgcagcatat agaaagtata cagaacagat tacaaatgag aagctggcta
      721 tggttaaagc ggaaccagat gttaaaaaat tagaagacca acttcaaggc ggtcaattag
      781 aagaggtgat tcttcaggct gaacatgaac taaatctggc aagaaaaatg agggaatgga
      841 aactatggga gccattagtg gaagagcctc ctgccgatca gtggaaatgg ccaatattaa
      901 acattggtgg gatatcaaat tctgcagtcg acggtaccag caagggcgag gagctgttca
      961 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1021 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ctcatctgca
     1081 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctcggc tacggcctgc
     1141 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1201 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1261 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1321 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     1381 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
     1441 acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     1501 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc aagctgagca
     1561 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     1621 tcactctcgg catggacgag ctgtacaagg gatccaccgg atctagataa ctgatcataa
     1681 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     1741 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     1801 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     1861 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     1921 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1981 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2041 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2101 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2161 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2221 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2281 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2341 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     2401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2461 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     2521 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2581 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2641 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2701 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2761 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2821 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     2881 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     2941 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3001 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3061 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3121 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3181 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3241 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3301 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3361 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     3421 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     3481 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     3541 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     3601 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     3661 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     3721 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3781 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3841 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     3901 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     3961 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4021 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4081 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4141 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4201 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     4681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     4741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     4801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4921 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4981 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     5041 atggagttcc g
//
