LOCUS       sequence-46256-e        8180 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8180)
  AUTHORS   Koivunen P, Lee S, Duncan CG, Lopez G, Lu G, Ramkissoon S, Losman 
            JA, Joensuu P, Bergmann U, Gross S, Travins J, Weiss S, Looper R, 
            Ligon KL, Verhaak RG, Yan H, Kaelin WG Jr
  TITLE     Transformation by the (R)-enantiomer of 2-hydroxyglutarate linked to
            EGLN activation.
  JOURNAL   Nature. 2012 Feb 15;483(7390):484-8. doi: 10.1038/nature10898.
  PUBMED    22343896
REFERENCE   2  (bases 1 to 8180)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "Nature"; date: "2012-02-15- 15"; volume: "483"; issue: "7390"; 
            pages: "484-8"
COMMENT     Sequence Label: FLAG-EglN1-pLenti6
FEATURES             Location/Qualifiers
     source          1..8180
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        3..229
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             230..410
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    457..582
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1075..1308
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     enhancer        1816..2119
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2120..2323
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2274..2294
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2320..2344
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2408..2431
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3768..3788)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3773..4102
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3953..4088
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4150..4197
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4216..4614
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4701..4934
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5006..5127
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5043..5062)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5097..5116
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5167..5302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5322..5341)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5323..5341)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5351..5368)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5351..5367)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5360..5382)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5509..5964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5596..5615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5806..5827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5990..6094
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6095..6163,6164..6955)
                     /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     complement(6313..6332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7126..7714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7615..7634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7868..7885
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8002..8023
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8038..8043,8044..8061,8062..8068)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8076..8092
                     /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)."
     primer_bind     8081..8103
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8100..8116
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8100..8116
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8135..8155
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8137..8155
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 aatgtagtct tatgcaatac tcttgtagtc ttgcaacatg gtaacgatga gttagcaaca
       61 tgccttacaa ggagagaaaa agcaccgtgc atgccgattg gtggaagtaa ggtggtacga
      121 tcgtgcctta ttaggaaggc aacagacggg tctgacatgg attggacgaa ccactgaatt
      181 gccgcattgc agagatattg tatttaagtg cctagctcga tacataaacg ggtctctctg
      241 gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc
      301 tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg
      361 taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca gtggcgcccg
      421 aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag gactcggctt
      481 gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc aaaaattttg
      541 actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa gcgggggaga
      601 attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa atataaatta
      661 aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc tggcctgtta
      721 gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct tcagacagga
      781 tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt gcatcaaagg
      841 atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca aaacaaaagt
      901 aagaccaccg cacagcaagc ggccgctgat cttcagacct ggaggaggag atatgaggga
      961 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc
     1021 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc
     1081 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct
     1141 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag
     1201 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca
     1261 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg
     1321 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa
     1381 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcgata
     1801 agcttgggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     1861 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     1921 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     1981 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     2041 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     2101 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     2161 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     2221 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     2281 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     2341 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gactctagcc
     2401 caccatggac tacaaagacg atgacgataa agggccctct agactcgaca tggccaatga
     2461 cagcggcggg cccggcgggc cgagcccgag cgagcgagac cggcagtact gcgagctgtg
     2521 cgggaagatg gagaacctgc tgcgctgcag ccgctgccgc agctccttct actgctgcaa
     2581 ggagcaccag cgtcaggact ggaagaagca caagctcgtg tgccagggca gcgagggcgc
     2641 cctcggccac ggagtgggcc cacaccagca ttccggcccc gcgccgccgg ctgcagtgcc
     2701 gccgcccagg gccggggccc gggagcccag gaaggcagcg gcgcgccggg acaacgcctc
     2761 cggggacgcg gccaagggaa aagtaaaggc caagcccccg gccgacccag cggcggccgc
     2821 gtcgccgtgt cgtgcggccg ccggcggcca gggctcggcg gtggctgccg aagccgagcc
     2881 cggcaaggag gagccgccgg cccgctcatc gctgttccag gagaaggcga acctgtaccc
     2941 cccaagcaac acgcccgggg atgcgctgag ccccggcggc ggcctgcggc ccaacgggca
     3001 gacgaagccc ctgccggcgc tgaagctggc gctcgagtac atcgtgccgt gcatgaacaa
     3061 gcacggcatc tgtgtggtgg acgacttcct cggcaaggag accggacagc agatcggcga
     3121 cgaggtgcgc gccctgcacg acaccgggaa gttcacggac gggcagctgg tcagccagaa
     3181 gagtgactcg tccaaggaca tccgaggcga taagatcacc tggatcgagg gcaaggagcc
     3241 cggctgcgaa accattgggc tgctcatgag cagcatggac gacctgatac gccactgtaa
     3301 cgggaagctg ggcagctaca aaatcaatgg ccggacgaaa gccatggttg cttgttatcc
     3361 gggcaatgga acgggttatg tacgtcatgt tgataatcca aatggagatg gaagatgtgt
     3421 gacatgtata tattatctta ataaagactg ggatgccaag gtaagtggag gtatacttcg
     3481 aatttttcca gaaggcaaag cccagtttgc tgacattgaa cccaaatttg atagactgct
     3541 gtttttctgg tctgaccgtc gcaaccctca tgaagtacaa ccagcatatg ctacaaggta
     3601 cgcaataact gtttggtatt ttgatgcaga tgagagagca cgagctaaag taaaatatct
     3661 aacaggtgaa aaaggtgtga gggttgaact caataaacct tcagattcgg tcggtaaaga
     3721 cgtcttctag gaattccggt tagtaatgag tttggaatta attctgtgga atgtgtgtca
     3781 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3841 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3901 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3961 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4021 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4081 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     4141 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     4201 gtgaggaact aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga
     4261 gcaacggcta caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct
     4321 ctctctagcg acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct
     4381 tgtgcagaac tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt
     4441 atcgtcgcga tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag
     4501 gtgcttctcg atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg
     4561 acggcagttg ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcacaa
     4621 ttcgagctcg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     4681 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagac aagatctgct
     4741 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     4801 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     4861 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     4921 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     4981 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     5041 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5101 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     5161 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5221 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5281 tgaggaggct tttttggagg cctagggacg tacccaattc gccctatagt gagtcgtatt
     5341 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5401 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5461 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     5521 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5581 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5641 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5701 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     5761 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5821 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     5881 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5941 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     6001 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     6061 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     6121 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     6181 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     6241 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     6301 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     6361 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     6421 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     6481 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     6541 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     6601 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     6661 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     6721 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     6781 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     6841 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     6901 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     6961 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     7021 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     7081 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     7141 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     7201 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     7261 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     7321 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     7381 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     7441 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     7501 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     7561 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     7621 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     7681 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     7741 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     7801 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     7861 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     7921 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     7981 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     8041 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     8101 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     8161 aaagctggag ctgcaagctt
//
