LOCUS       sequence-237964-        7047 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 7047)
  AUTHORS   Zhang G, Yanamala N, Lathrop KL, Zhang L, Klein-Seetharaman J, 
            Srinivas H
  TITLE     Ligand-independent antiapoptotic function of estrogen receptor-beta 
            in lung cancer cells.
  JOURNAL   Mol Endocrinol. 2010 Sep;24(9):1737-47. Epub 2010 Jul 21.
  PUBMED    20660297
REFERENCE   2  (bases 1 to 7047)
  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: "Mol 
            Endocrinol."; date: "2010-09"; volume: "24(9)"; pages: "1737-47. 
            Epub 2010 Jul 21"
COMMENT     Sequence Label: pcDNA Flag ERbeta
FEATURES             Location/Qualifiers
     source          1..7047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     865..884
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        865..883
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             941..1006
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2640..2657)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2646..2870
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2916..3344
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3003..3022)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3213..3234
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3353..3373)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3358..3687
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3538..3673
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3600..3619
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3754..4548
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3808..3827)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4418..4437
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4722..4843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4759..4778)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4813..4832
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4892..4908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4892..4908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4905..4927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4916..4932
                     /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)."
     promoter        complement(join(4940..4946,4947..4964,4965..4970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4985..5006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5123..5140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5294..5882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5374..5393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6053..6844,6845..6913))
                     /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     6676..6695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6914..7018)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
      841 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
      901 gcgtttaaac ttaagcttgg taccgagctc ggatcctatg gactacaaag accatgacgg
      961 tgattataaa gatcatgaca tcgattacaa ggatgacgat gacaagctta tggatataaa
     1021 aaactcacca tctagcctta attctccttc ctcctacaac tgcagtcaat ccatcttacc
     1081 cctggagcac ggctccatat acataccttc ctcctatgta gacagccacc atgaatatcc
     1141 agccatgaca ttctatagcc ctgctgtgat gaattacagc attcccagca atgtcactaa
     1201 cttggaaggt gggcctggtc ggcagaccac aagcccaaat gtgttgtggc caacacctgg
     1261 gcacctttct cctttagtgg tccatcgcca gttatcacat ctgtatgcgg aacctcaaaa
     1321 gagtccctgg tgtgaagcaa gatcgctaga acacacctta cctgtaaaca gagagacact
     1381 gaaaaggaag gttagtggga accgttgcgc cagccctgtt actggtccag gttcaaagag
     1441 ggatgctcac ttctgcgctg tctgcagcga ttacgcatcg ggatatcact atggagtctg
     1501 gtcgtgtgaa ggatgtaagg ccttttttaa aagaagcatt caaggacata atgattatat
     1561 ttgtccagct acaaatcagt gtacaatcga taaaaaccgg cgcaagagct gccaggcctg
     1621 ccgacttcgg aagtgttacg aagtgggaat ggtgaagtgt ggctcccgga gagagagatg
     1681 tgggtaccgc cttgtgcgga gacagagaag tgccgacgag cagctgcact gtgccggcaa
     1741 ggccaagaga agtggcggcc acgcgccccg agtgcgggag ctgctgctgg acgccctgag
     1801 ccccgagcag ctagtgctca ccctcctgga ggctgagccg ccccatgtgc tgatcagccg
     1861 ccccagtgcg cccttcaccg aggcctccat gatgatgtcc ctgaccaagt tggccgacaa
     1921 ggagttggta cacatgatca gctgggccaa gaagattccc ggctttgtgg agctcagcct
     1981 gttcgaccaa gtgcggctct tggagagctg ttggatggag gtgttaatga tggggctgat
     2041 gtggcgctca attgaccacc ccggcaagct catctttgct ccagatcttg ttctggacag
     2101 ggatgagggg aaatgcgtag aaggaattct ggaaatcttt gacatgctcc tggcaactac
     2161 ttcaaggttt cgagagttaa aactccaaca caaagaatat ctctgtgtca aggccatgat
     2221 cctgctcaat tccagtatgt accctctggt cacagcgacc caggatgctg acagcagccg
     2281 gaagctggct cacttgctga acgccgtgac cgatgctttg gtttgggtga ttgccaagag
     2341 cggcatctcc tcccagcagc aatccatgcg cctggctaac ctcctgatgc tcctgtccca
     2401 cgtcaggcat gcgagtaaca agggcatgga acatctgctc aacatgaagt gcaaaaatgt
     2461 ggtcccagtg tatgacctgc tgctggagat gctgaatgcc cacgtgcttc gcgggtgcaa
     2521 gtcctccatc acggggtccg agtgcagccc ggcagaggac agtaaaagca aagagggctc
     2581 ccagaaccca cagtctcagt gactcgagtc tagagggccc gtttaaaccc gctgatcagc
     2641 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt
     2701 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca
     2761 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga
     2821 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc
     2881 ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag
     2941 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     3001 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3061 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3121 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3181 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3241 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     3301 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg
     3361 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3421 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3481 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3541 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3601 atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc
     3661 ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga
     3721 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3781 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3841 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3901 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     3961 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     4021 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     4081 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     4141 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4201 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4261 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     4321 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4381 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4441 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4501 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4561 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4621 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4681 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4741 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4801 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4861 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4921 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4981 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5041 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5101 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5161 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5221 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5281 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5341 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5401 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5461 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5521 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5581 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5641 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5701 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5761 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5821 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5881 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5941 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6001 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6061 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6121 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6181 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6241 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6301 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6361 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6421 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6481 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6541 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6601 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6661 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6721 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6781 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6841 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6901 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6961 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7021 catttccccg aaaagtgcca cctgacg
//
