LOCUS       sequence-46258-e        7619 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 7619)
  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 7619)
  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-EglN3-pLenti6
FEATURES             Location/Qualifiers
     source          1..7619
                     /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(3207..3227)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3212..3541
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3392..3527
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3589..3636
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3655..4053
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4140..4373
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4445..4566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4482..4501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4536..4555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4606..4741
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4761..4780)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4762..4780)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4790..4807)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4790..4806)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4799..4821)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4948..5403
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5035..5054)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5245..5266
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5429..5533
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5534..5602,5603..6394)
                     /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(5752..5771)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6565..7153
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7054..7073
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7307..7324
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7441..7462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7477..7482,7483..7500,7501..7507)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7515..7531
                     /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     7520..7542
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7539..7555
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7539..7555
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7574..7594
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7576..7594
                     /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 tgcccctggg
     2461 acacatcatg aggctggacc tggagaaaat tgccctggag tacatcgtgc cctgtctgca
     2521 cgaggtgggc ttctgctacc tggacaactt cctgggcgag gtggtgggcg actgcgtcct
     2581 ggagcgcgtc aagcagctgc actgcaccgg ggccctgcgg gacggccagc tggcggggcc
     2641 gcgcgccggc gtctccaagc gacacctgcg gggcgaccag atcacgtgga tcgggggcaa
     2701 cgaggagggc tgcgaggcca tcagcttcct cctgtccctc atcgacaggc tggtcctcta
     2761 ctgcgggagc cggctgggca aatactacgt caaggagagg tctaaggcaa tggtggcttg
     2821 ctatccggga aatggaacag gttatgttcg ccacgtggac aaccccaacg gtgatggtcg
     2881 ctgcatcacc tgcatctact atctgaacaa gaattgggat gccaagctac atggtgggat
     2941 cctgcggata tttccagagg ggaaatcatt catagcagat gtggagccca tttttgacag
     3001 actcctgttc ttctggtcag atcgtaggaa cccacacgaa gtgcagccct cttacgcaac
     3061 cagatatgct atgactgtct ggtactttga tgctgaagaa agggcagaag ccaaaaagaa
     3121 attcaggaat ttaactagga aaactgaatc tgccctcact gaagactgag aattccggtt
     3181 agtaatgagt ttggaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3241 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3301 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3361 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3421 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3481 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3541 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     3601 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     3661 aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc
     3721 atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc
     3781 ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg
     3841 ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag
     3901 aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct
     3961 gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa
     4021 ttgctgccct ctggttatgt gtgggagggc taagcacaat tcgagctcgg tacctttaag
     4081 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     4141 ggaagggcta attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct
     4201 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     4261 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     4321 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     4381 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     4441 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4501 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4561 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact
     4621 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4681 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4741 ctagggacgt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     4801 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     4861 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     4921 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     4981 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     5041 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     5101 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     5161 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     5221 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     5281 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     5341 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     5401 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5461 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5521 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5581 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5641 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5701 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5761 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5821 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5881 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5941 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6001 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6061 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6121 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6181 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6241 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6301 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6361 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6421 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6481 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6541 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6601 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6661 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     6721 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6781 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6841 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6901 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6961 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7021 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7081 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7141 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7201 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     7261 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7321 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     7381 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     7441 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     7501 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     7561 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tgcaagctt
//
