LOCUS       sequence-10381-e        6405 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 6405)
  AUTHORS   Zhang Q, Gu J, Li L, Liu J, Luo B, Cheung HW, Boehm JS, Ni M, Geisen
            C, Root DE, Polyak K, Brown M, Richardson AL, Hahn WC, Kaelin WG, 
            Bommi-Reddy A
  TITLE     Control of Cyclin D1 and Breast Tumorigenesis by the EglN2 Prolyl 
            Hydroxylase.
  JOURNAL   Cancer Cell. 2009 Nov 6. 16(5):413-24.
  PUBMED    19878873
REFERENCE   2  (bases 1 to 6405)
  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"; journalName: "Cancer 
            Cell. 2009 Nov 6. 16(5):413-24."
COMMENT     Sequence Label: FLAG-EglN2 wt-pBabe-Puro
FEATURES             Location/Qualifiers
     source          1..6405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             934..1350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1318..1334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1370..1393
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2235..2244
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2644..2664)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2649..2978
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2829..2964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2988..3587
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     primer_bind     complement(2988..3007)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3484..3504
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3686..4279
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4480..4497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4651..5239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4731..4750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5417..6208,6209..6277))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6040..6059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6278..6382)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt
      901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
      961 ttgaccttag atcactggaa agatgtcgag cggctcgctc acaaccagtc ggtagatgtc
     1021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     1081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     1141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     1201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttctt
     1261 ccatccgcgc cgtctctccc ccttgaacct cctctttcga ccccgcctca atcctccctt
     1321 tatccagccc tcactccttc tctaggcgcc ggccggatcc cggaggatgg actacaaaga
     1381 cgatgacgac aaggacagcc cgtgccagcc gcagccccta agtcaggctc tccctcagtt
     1441 accagggtct tcgtcagagc ccttggagcc tgagcctggc cgggccagga tgggagtgga
     1501 gagttacctg ccctgtcccc tgctcccctc ctaccactgt ccaggagtgc ctagtgaggc
     1561 ctcggcaggg agtgggaccc ccagagccac agccacctct accactgcca gccctcttcg
     1621 ggacggtttt ggcgggcagg atggtggtga gctgcggccg ctgcagagtg aaggcgctgc
     1681 agcgctggtc accaaggggt gccagcgatt ggcagcccag ggcgcacggc ctgaggcccc
     1741 caaacggaaa tgggccgagg atggtgggga tgccccttca cccagcaaac ggccctgggc
     1801 caggcaagag aaccaggagg cagagcggga gggtggcatg agctgcagct gcagcagtgg
     1861 cagtggtgag gccagtgctg ggctgatgga ggaggcgctg ccctctgcgc ccgagcgcct
     1921 ggccctggac tatatcgtgc cctgcatgcg gtactacggc atctgcgtca aggacagctt
     1981 cctgggggca gcactgggcg gtcgcgtgct ggccgaggtg gaggccctca aacggggtgg
     2041 gcgcctgcga gacgggcagc tagtgagcca gagggcgatc ccgccgcgca gcatccgtgg
     2101 ggaccagatt gcctgggtgg aaggccatga accaggctgt cgaagcattg gtgccctcat
     2161 ggcccatgtg gacgccgtca tccgccactg cgcagggcgg ctgggcagct atgtcatcaa
     2221 cgggcgcacc aaggccatgg tggcgtgtta cccaggcaac gggctcgggt acgtaaggca
     2281 cgttgacaat ccccacggcg atgggcgctg catcacctgt atctattacc tgaatcagaa
     2341 ctgggacgtt aaggtgcatg gcggcctgct gcagatcttc cctgagggcc ggcccgtggt
     2401 agccaacatc gagccactct ttgaccggtt gctcattttc tggtctgacc ggcggaaccc
     2461 ccacgaggtg aagccagcct atgccaccag gtacgccatc actgtctggt attttgatgc
     2521 caaggagcgg gcagcagcca aagacaagta tcagctagca tcaggacaga aaggtgtcca
     2581 agtacctgta tcacagccgc ctacgcccac ctaggaattc gccagcacag tggtcgaccc
     2641 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2701 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2821 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2881 taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt
     2941 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttaccatg accgagtaca
     3001 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
     3061 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
     3121 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
     3181 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
     3241 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
     3301 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
     3361 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
     3421 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
     3481 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgagtgcc
     3541 cgaaggaccg cgcgacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
     3601 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcgataaaa taaaagattt
     3661 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     3721 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     3781 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     3841 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     3901 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     3961 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4021 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4081 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     4141 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4201 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4261 tcagcggggg tctttcattt ccgacttgtg gtctcgctgc cttgggaggg tctcctctga
     4321 gtgattgact acccgtcagc gggggtcttc acatgcagca tgtatcaaaa ttaatttggt
     4381 tttttttctt aagtatttac attaaatggc catagttgca ttaatgaatc ggccaacgcg
     4441 cggggagagg cggtttgcgt attggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4501 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4561 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4621 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4681 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4741 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4801 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4861 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4921 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4981 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5041 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     5101 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5161 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5221 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5281 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5341 gatcctttta aattaaaaat gaagtttgcg gccgcaaatc aatctaaagt atatatgagt
     5401 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5461 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5521 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5581 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5641 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5701 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5761 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5821 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5881 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5941 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6001 tgcggcgacc gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca
     6061 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6121 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6181 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6241 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6301 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6361 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccac
//
