LOCUS       sequence-288038-        6544 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 6544)
  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 6544)
  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: pGEX 4T1 ERbeta
FEATURES             Location/Qualifiers
     source          1..6544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(83..88,89..104,105..111)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    119..135
                     /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)."
     CDS             158..811
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     769..791
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             818..835
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2494..2516)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2661..2679)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2747..2851
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2852..2920,2921..3712)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3070..3089)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3883..4471
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4372..4391
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4625..4642
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4715..4792
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4793..5875
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4812..4831)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5924..5929,5930..5947,5948..5954)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5962..5978
                     /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     5986..6002
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5986..6002
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5998..6525
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6018..6035)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6018..6034)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6027..6049)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 cgctcaaggc gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg
       61 gcaaatattc tgaaatgagc tgttgacaat taatcatcgg ctcgtataat gtgtggaatt
      121 gtgagcggat aacaatttca cacaggaaac agtattcatg tcccctatac taggttattg
      181 gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata
      241 tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt
      301 gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc
      361 tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga
      421 gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag
      481 aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga
      541 aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt
      601 aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat
      661 gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca
      721 aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc
      781 cacgtttggt ggtggcgacc atcctccaaa atcggatctg gttccgcgtg gatccatgga
      841 tataaaaaac tcaccatcta gccttaattc tccttcctcc tacaactgca gtcaatccat
      901 cttacccctg gagcacggct ccatatacat accttcctcc tatgtagaca gccaccatga
      961 atatccagcc atgacattct atagccctgc tgtgatgaat tacagcattc ccagcaatgt
     1021 cactaacttg gaaggtgggc ctggtcggca gaccacaagc ccaaatgtgt tgtggccaac
     1081 acctgggcac ctttctcctt tagtggtcca tcgccagtta tcacatctgt atgcggaacc
     1141 tcaaaagagt ccctggtgtg aagcaagatc gctagaacac accttacctg taaacagaga
     1201 gacactgaaa aggaaggtta gtgggaaccg ttgcgccagc cctgttactg gtccaggttc
     1261 aaagagggat gctcacttct gcgctgtctg cagcgattac gcatcgggat atcactatgg
     1321 agtctggtcg tgtgaaggat gtaaggcctt ttttaaaaga agcattcaag gacataatga
     1381 ttatatttgt ccagctacaa atcagtgtac aatcgataaa aaccggcgca agagctgcca
     1441 ggcctgccga cttcggaagt gttacgaagt gggaatggtg aagtgtggct cccggagaga
     1501 gagatgtggg taccgccttg tgcggagaca gagaagtgcc gacgagcagc tgcactgtgc
     1561 cggcaaggcc aagagaagtg gcggccacgc gccccgagtg cgggagctgc tgctggacgc
     1621 cctgagcccc gagcagctag tgctcaccct cctggaggct gagccgcccc atgtgctgat
     1681 cagccgcccc agtgcgccct tcaccgaggc ctccatgatg atgtccctga ccaagttggc
     1741 cgacaaggag ttggtacaca tgatcagctg ggccaagaag attcccggct ttgtggagct
     1801 cagcctgttc gaccaagtgc ggctcttgga gagctgttgg atggaggtgt taatgatggg
     1861 gctgatgtgg cgctcaattg accaccccgg caagctcatc tttgctccag atcttgttct
     1921 ggacagggat gaggggaaat gcgtagaagg aattctggaa atctttgaca tgctcctggc
     1981 aactacttca aggtttcgag agttaaaact ccaacacaaa gaatatctct gtgtcaaggc
     2041 catgatcctg ctcaattcca gtatgtaccc tctggtcaca gcgacccagg atgctgacag
     2101 cagccggaag ctggctcact tgctgaacgc cgtgaccgat gctttggttt gggtgattgc
     2161 caagagcggc atctcctccc agcagcaatc catgcgcctg gctaacctcc tgatgctcct
     2221 gtcccacgtc aggcatgcga gtaacaaggg catggaacat ctgctcaaca tgaagtgcaa
     2281 aaatgtggtc ccagtgtatg acctgctgct ggagatgctg aatgcccacg tgcttcgcgg
     2341 gtgcaagtcc tccatcacgg ggtccgagtg cagcccggca gaggacagta aaagcaaaga
     2401 gggctcccag aacccacagt ctcagtgact cgagcggccg catcgtgact gactgacgat
     2461 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     2521 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     2581 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     2641 taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     2701 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2761 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2821 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2881 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2941 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     3001 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     3061 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     3121 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     3181 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     3241 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     3301 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3361 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     3421 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3481 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3541 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3601 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3661 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3721 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3781 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     3841 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     3901 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     3961 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     4021 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     4081 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     4141 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     4201 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     4261 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     4321 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     4381 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     4441 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     4501 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     4561 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     4621 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
     4681 gcatctgtgc ggtatttcac accgcataaa ttccgacacc atcgaatggc gcaaaacctt
     4741 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
     4801 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
     4861 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
     4921 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
     4981 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
     5041 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
     5101 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
     5161 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
     5221 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
     5281 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
     5341 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
     5401 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
     5461 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
     5521 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
     5581 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
     5641 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
     5701 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
     5761 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     5821 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     5881 acgcaattaa tgtaagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     5941 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     6001 accatgatta cggattcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     6061 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     6121 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt
     6181 gcctggtttc cggcaccaga agcggtgccg gaaagctggc tggagtgcga tcttcctgag
     6241 gccgatactg tcgtcgtccc ctcaaactgg cagatgcacg gttacgatgc gcccatctac
     6301 accaacgtga cctatcccat tacggtcaat ccgccgtttg ttcccacgga gaatccgacg
     6361 ggttgttact cgctcacatt taatgttgat gaaagctggc tacaggaagg ccagacgcga
     6421 attatttttg atggcgttgg aattagctta tcgactgcac ggtgcaccaa tgcttctggc
     6481 gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc
     6541 gtgt
//
