LOCUS       sequence-46257-e        8122 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 8122)
  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 8122)
  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-EglN2-pLenti6
FEATURES             Location/Qualifiers
     source          1..8122
                     /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"
     regulatory      2398..2407
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2407..2430
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      3293..3302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3710..3730)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3715..4044
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3895..4030
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4092..4139
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4158..4556
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4643..4876
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4948..5069
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4985..5004)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5039..5058
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5109..5244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5264..5283)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5265..5283)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5293..5310)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5293..5309)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5302..5324)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5451..5906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5538..5557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5748..5769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5932..6036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6037..6105,6106..6897)
                     /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(6255..6274)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7068..7656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7557..7576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7810..7827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7944..7965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7980..7985,7986..8003,8004..8010)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8018..8034
                     /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     8023..8045
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8042..8058
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8042..8058
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     8077..8097
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        8079..8097
                     /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 accatggact acaaagacga tgacgataaa gggccctcta gactcgacat ggacagcccg
     2461 tgccagccgc agcccctaag tcaggctctc cctcagttac cagggtcttc gtcagagccc
     2521 ttggagcctg agcctggccg ggccaggatg ggagtggaga gttacctgcc ctgtcccctg
     2581 ctcccctcct accactgtcc aggagtgcct agtgaggcct cggcagggag tgggaccccc
     2641 agagccacag ccacctctac cactgccagc cctcttcggg acggttttgg cgggcaggat
     2701 ggtggtgagc tgcggccgct gcagagtgaa ggcgctgcag cgctggtcac caaggggtgc
     2761 cagcgattgg cagcccaggg cgcacggcct gaggccccca aacggaaatg ggccgaggat
     2821 ggtggggatg ccccttcacc cagcaaacgg ccctgggcca ggcaagagaa ccaggaggca
     2881 gagcgggagg gtggcatgag ctgcagctgc agcagtggca gtggtgaggc cagtgctggg
     2941 ctgatggagg aggcgctgcc ctctgcgccc gagcgcctgg ccctggacta tatcgtgccc
     3001 tgcatgcggt actacggcat ctgcgtcaag gacagcttcc tgggggcagc actgggcggt
     3061 cgcgtgctgg ccgaggtgga ggccctcaaa cggggtgggc gcctgcgaga cgggcagcta
     3121 gtgagccaga gggcgatccc gccgcgcagc atccgtgggg accagattgc ctgggtggaa
     3181 ggccatgaac caggctgtcg aagcattggt gccctcatgg cccatgtgga cgccgtcatc
     3241 cgccactgcg cagggcggct gggcagctat gtcatcaacg ggcgcaccaa ggccatggtg
     3301 gcgtgttacc caggcaacgg gctcgggtac gtaaggcacg ttgacaatcc ccacggcgat
     3361 gggcgctgca tcacctgtat ctattacctg aatcagaact gggacgttaa ggtgcatggc
     3421 ggcctgctgc agatcttccc tgagggccgg cccgtggtag ccaacatcga gccactcttt
     3481 gaccggttgc tcattttctg gtctgaccgg cggaaccccc acgaggtgaa gccagcctat
     3541 gccaccaggt acgccatcac tgtctggtat tttgatgcca aggagcgggc agcagccaaa
     3601 gacaagtatc agctagcatc aggacagaaa ggtgtccaag tacctgtatc acagccgcct
     3661 acgcccacct aggaattccg gttagtaatg agtttggaat taattctgtg gaatgtgtgt
     3721 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3781 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     3841 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     3901 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     3961 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4021 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4081 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     4141 aggtgaggaa ctaaaccatg gccaagcctt tgtctcaaga agaatccacc ctcattgaaa
     4201 gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc gccagcgcag
     4261 ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt actgggggac
     4321 cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc aacctgactt
     4381 gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga cggtgccgac
     4441 aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt gatggacagc
     4501 cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag ggctaagcac
     4561 aattcgagct cggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac
     4621 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatctg
     4681 ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga gctctctggc
     4741 taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct tcaagtagtg
     4801 tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt ttagtcagtg
     4861 tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt ataacttgca
     4921 aagaaatgaa tatcagagag tgagaggaac ttgtttattg cagcttataa tggttacaaa
     4981 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5041 ggtttgtcca aactcatcaa tgtatcttat catgtctggc tctagctatc ccgcccctaa
     5101 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5161 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     5221 agtgaggagg cttttttgga ggcctaggga cgtacccaat tcgccctata gtgagtcgta
     5281 ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5341 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5401 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     5461 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5521 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5581 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5641 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5701 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5761 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5821 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5881 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     5941 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6001 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6061 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6121 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6181 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6241 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6301 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6361 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6421 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6481 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6541 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6601 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6661 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6721 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6781 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6841 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6901 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6961 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7021 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7081 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7141 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7201 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7261 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7321 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7381 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7441 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7501 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7561 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7621 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7681 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7741 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7801 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7861 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7921 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7981 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     8041 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     8101 caaaagctgg agctgcaagc tt
//
