LOCUS       sequence-46255-e        7611 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 7611)
  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 7611)
  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-Venus-pLenti6
FEATURES             Location/Qualifiers
     source          1..7611
                     /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"
     CDS             join(2444..2446,2447..2449,2450..3157)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELY"
     primer_bind     complement(2489..2510)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2750..2769)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3097..3118
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3199..3219)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3204..3533
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3384..3519
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3581..3628
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3647..4045
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             4132..4365
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    4437..4558
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4474..4493)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4528..4547
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4598..4733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4753..4772)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4754..4772)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4782..4799)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4782..4798)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4791..4813)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4940..5395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5027..5046)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5237..5258
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5421..5525
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5526..5594,5595..6386)
                     /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(5744..5763)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6557..7145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7046..7065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7299..7316
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7433..7454
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7469..7474,7475..7492,7493..7499)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7507..7523
                     /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     7512..7534
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7531..7547
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7531..7547
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7566..7586
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7568..7586
                     /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 agaatggtga gcaagggcga
     2461 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2521 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2581 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     2641 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2701 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2761 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2821 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2881 caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     2941 caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa
     3001 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc
     3061 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3121 cgccgccggg atcactctcg gcatggacga gctgtacata aggatcccgg ttagtaatga
     3181 gtttggaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3361 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3481 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3541 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     3601 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt
     3661 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     3721 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     3781 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     3841 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     3901 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     3961 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     4021 ctctggttat gtgtgggagg gctaagcaca attcgagctc ggtaccttta agaccaatga
     4081 cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc
     4141 taattcactc ccaacgaaga caagatctgc tttttgcttg tactgggtct ctctggttag
     4201 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
     4261 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
     4321 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
     4381 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggaact
     4441 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4501 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4561 atgtctggct ctagctatcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     4621 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     4681 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctagggac
     4741 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     4801 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4861 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4921 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4981 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     5041 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     5101 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     5161 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     5221 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     5281 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     5341 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     5401 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5461 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5521 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5581 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5641 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5701 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5761 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5821 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5881 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5941 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6001 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6061 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6121 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6181 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6241 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6301 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6361 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6421 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6481 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6541 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6601 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6661 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6721 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6781 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6841 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6901 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6961 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7021 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7081 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7141 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7201 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7261 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7321 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7381 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7441 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7501 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     7561 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct t
//
