LOCUS       Exported                6194 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    HA-VHL R167W-pRc/CMV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6194)
  AUTHORS   Kibel A, Iliopoulos O, DeCaprio JA, Kaelin WG
  TITLE     Binding of the von Hippel-Lindau tumor suppressor protein to Elongin
            B and C.
  JOURNAL   Science. 1995 Sep 8. 269(5229):1444-6.
  PUBMED    7660130
REFERENCE   2  (bases 1 to 6194)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6194
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             904..930
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1651..1669)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1652..1669)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1689..1706)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1695..1919
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1965..2478
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2052..2071)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2262..2283
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2501..2521)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2506..2836
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2687..2822
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2749..2768
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2903..3697
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2957..2976)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3567..3586
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3871..4005
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3908..3927)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3962..3981
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4037..4053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4037..4053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4050..4072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4061..4077
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4085..4115)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4130..4151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4268..4285)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4439..5027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4519..4538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5198..6058)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5821..5840
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6059..6163)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
      901 gcctacccct acgacgtgcc cgactacgcc tccctcggat ccgccaccat gccccggagg
      961 gcggagaact gggacgaggc cgaggtaggc gcggaggagg caggcgtcga agagtacggc
     1021 cctgaagaag acggcgggga ggagtcgggc gccgaggagt ccggcccgga agagtccggc
     1081 ccggaggaac tgggcgccga ggaggagatg gaggccgggc ggccgcggcc cgtgctgcgc
     1141 tcggtgaact cgcgcgagcc ctcccaggtc atcttctgca atcgcagtcc gcgcgtcgtg
     1201 ctgcccgtat ggctcaactt cgacggcgag ccgcagccct acccaacgct gccgcctggc
     1261 acgggccgcc gcatccacag ctaccgaggt cacctttggc tcttcagaga tgcagggaca
     1321 cacgatgggc ttctggttaa ccaaactgaa ttatttgtgc catctctcaa tgttgacgga
     1381 cagcctattt ttgccaatat cacactgcca gtgtatactc tgaaagagcg atgcctccag
     1441 gttgtctgga gcctagtcaa gcctgagaat tacaggagac tggacatcgt caggtcgctc
     1501 tacgaagatc tggaagacca cccaaatgtg cagaaagacc tggagcggct gacacaggag
     1561 cgcattgcac atcaacggat gggagattga attctgcaga tatccatcac actggcggcc
     1621 gctcgagcat gcatctctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     1681 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1741 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1801 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1861 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     1921 ttctgaggcg gaaagaacca gctggggctc gaggggggat ccccacgcgc cctgtagcgg
     1981 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2041 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2101 ccgtcaagct ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct
     2161 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2221 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2281 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat
     2341 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2401 aatattaacg tttacaattt aaatatttgc ttatacaatc ttcctgtttt tggggctttt
     2461 ctgattatca accggggtgg gtaccgagct cgaattctgt ggaatgtgtg tcagttaggg
     2521 tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc atctcaatta
     2581 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2641 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2701 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2761 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2821 cctaggcttt tgcaaaaagc tcccgggagc ttggatatcc attttcggat ctgatcaaga
     2881 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     2941 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3001 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3061 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3121 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3181 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3241 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3301 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3361 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3421 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3481 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3541 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3601 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3661 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3721 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3781 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3841 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3901 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3961 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatcccgtc gacctcgaga
     4021 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4081 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     4141 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     4201 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4261 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4321 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4381 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4621 tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4861 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5161 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5221 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5281 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5341 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5401 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5461 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5521 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5581 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5641 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5701 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5761 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5821 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5881 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5941 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6001 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6061 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6121 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6181 tgccacctga cgtc
//
