LOCUS       Exported                7937 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    HA-HIF1alpha-pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7937)
  AUTHORS   Kondo K, Klco J, Nakamura E, Lechpammer M, Kaelin WG
  TITLE     Inhibition of HIF is necessary for tumor suppression by the von 
            Hippel-Lindau protein.
  JOURNAL   Cancer Cell. 2002 Apr . 1(3):237-46.
  PUBMED    12086860
REFERENCE   2  (bases 1 to 7937)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7937
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             906..932
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      945..954
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     promoter        complement(3492..3510)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3493..3510)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3530..3547)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3536..3760
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3806..4234
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3893..3912)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4103..4124
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4243..4263)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4248..4577
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4428..4563
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4490..4509
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4644..5438
                     /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(4698..4717)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5308..5327
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5612..5733
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5649..5668)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5703..5722
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5782..5798)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5782..5798)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5795..5817)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5806..5822
                     /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(5830..5860)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5875..5896
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6013..6030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6184..6772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6264..6283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6943..7803)
                     /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     7566..7585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7804..7908)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagcttacca
      901 tggcctaccc ctacgacgtg cccgactacg cctccctcgg atccgccacc atggagggcg
      961 ccggcggcgc gaacgacaag aaaaagataa gttctgaacg tcgaaaagaa aagtctcgag
     1021 atgcagccag atctcggcga agtaaagaat ctgaagtttt ttatgagctt gctcatcagt
     1081 tgccacttcc acataatgtg agttcgcatc ttgataaggc ctctgtgatg aggcttacca
     1141 tcagctattt gcgtgtgagg aaacttctgg atgctggtga tttggatatt gaagatgaca
     1201 tgaaagcaca gatgaattgc ttttatttga aagccttgga tggttttgtt atggttctca
     1261 cagatgatgg tgacatgatt tacatttctg ataatgtgaa caaatacatg ggattaactc
     1321 agtttgaact aactggacac agtgtgtttg attttactca tccatgtgac catgaggaaa
     1381 tgagagaaat gcttacacac agaaatggcc ttgtgaaaaa gggtaaagaa caaaacacac
     1441 agcgaagctt ttttctcaga atgaagtgta ccctaactag ccgaggaaga actatgaaca
     1501 taaagtctgc aacatggaag gtattgcact gcacaggcca cattcacgta tatgatacca
     1561 acagtaacca acctcagtgt gggtataaga aaccacctat gacctgcttg gtgctgattt
     1621 gtgaacccat tcctcaccca tcaaatattg aaattccttt agatagcaag actttcctca
     1681 gtcgacacag cctggatatg aaattttctt attgtgatga aagaattacc gaattgatgg
     1741 gatatgagcc agaagaactt ttaggccgct caatttatga atattatcat gctttggact
     1801 ctgatcatct gaccaaaact catcatgata tgtttactaa aggacaagtc accacaggac
     1861 agtacaggat gcttgccaaa agaggtggat atgtctgggt tgaaactcaa gcaactgtca
     1921 tatataacac caagaattct caaccacagt gcattgtatg tgtgaattac gttgtgagtg
     1981 gtattattca gcacgacttg attttctccc ttcaacaaac agaatgtgtc cttaaaccgg
     2041 ttgaatcttc agatatgaaa atgactcagc tattcaccaa agttgaatca gaagatacaa
     2101 gtagcctctt tgacaaactt aagaaggaac ctgatgcttt aactttgctg gccccagccg
     2161 ctggagacac aatcatatct ttagattttg gcagcaacga cacagaaact gatgaccagc
     2221 aacttgagga agtaccatta tataatgatg taatgctccc ctcacccaac gaaaaattac
     2281 agaatataaa tttggcaatg tctccattac ccaccgctga aacgccaaag ccacttcgaa
     2341 gtagtgctga ccctgcactc aatcaagaag ttgcattaaa attagaacca aatccagagt
     2401 cactggaact ttcttttacc atgccccaga ttcaggatca gacacctagt ccttccgatg
     2461 gaagcactag acaaagttca cctgagccta atagtcccag tgaatattgt ttttatgtgg
     2521 atagtgatat ggtcaatgaa ttcaagttgg aattggtaga aaaacttttt gctgaagaca
     2581 cagaagcaaa gaacccattt tctactcagg acacagattt agacttggag atgttagctc
     2641 cctatatccc aatggatgat gacttccagt tacgttcctt cgatcagttg tcaccattag
     2701 aaagcagttc cgcaagccct gaaagcgcaa gtcctcaaag cacagttaca gtattccagc
     2761 agactcaaat acaagaacct actgctaatg ccaccactac cactgccacc actgatgaat
     2821 taaaaacagt gacaaaagac cgtatggaag acattaaaat attgattgca tctccatctc
     2881 ctacccacat acataaagaa actactagtg ccacatcatc accatataga gatactcaaa
     2941 gtcggacagc ctcaccaaac agagcaggaa aaggagtcat agaacagaca gaaaaatctc
     3001 atccaagaag ccctaacgtg ttatctgtcg ctttgagtca aagaactaca gttcctgagg
     3061 aagaactaaa tccaaagata ctagctttgc agaatgctca gagaaagcga aaaatggaac
     3121 atgatggttc actttttcaa gcagtaggaa ttggaacatt attacagcag ccagacgatc
     3181 atgcagctac tacatcactt tcttggaaac gtgtaaaagg atgcaaatct agtgaacaga
     3241 atggaatgga gcaaaagaca attattttaa taccctctga tttagcatgt agactgctgg
     3301 ggcaatcaat ggatgaaagt ggattaccac agctgaccag ttatgattgt gaagttaatg
     3361 ctcctataca aggcagcaga aacctactgc agggtgaaga attactcaga gctttggatc
     3421 aagttaactg acaattctgc agatatccat cacactggcg gccgctcgag catgcatcta
     3481 gagggcccta ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg
     3541 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     3601 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     3661 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     3721 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     3781 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3841 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3901 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3961 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     4021 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     4081 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     4141 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     4201 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     4261 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4321 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4381 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4441 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4501 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4561 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     4621 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4681 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4741 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4801 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4861 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4921 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4981 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     5041 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     5101 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     5161 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     5221 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     5281 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     5341 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5401 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5461 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5521 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5581 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     5641 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5701 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     5761 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5821 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5881 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5941 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6001 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6061 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     6661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7921 aaaagtgcca cctgacg
//
