LOCUS       sequence-234694-        6616 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of SRF as a GST fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  AUTHORS   Herring BP, Kriegel AM, Hoggatt AM
  TITLE     Identification of Barx2b, a serum response factor-associated 
            homeodomain protein.
  JOURNAL   J Biol Chem. 2001 Apr 27;276(17):14482-9. doi: 
            10.1074/jbc.M011585200. Epub 2001 Jan 29.
  PUBMED    11278942
REFERENCE   2  (bases 1 to 6616)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M011585200"; journalName: "J Biol Chem"; date: 
            "2001-04-27- 27"; volume: "276"; issue: "17"; pages: "14482-9"
COMMENT     Sequence Label: SRF WT pGEX4T
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(51..56,57..72,73..79)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    87..103
                     /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             126..779
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     737..759
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             786..803
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2534..2556)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2701..2719)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2787..2891
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2892..2960,2961..3752)
                     /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(3110..3129)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3923..4511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4412..4431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4665..4682
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4755..4832
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4833..5915
                     /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(4852..4871)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5964..5969,5970..5987,5988..5994)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6002..6018
                     /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     6026..6042
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6026..6042
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6038..6565
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(6058..6075)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6058..6074)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6067..6089)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
       61 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
      121 tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac
      181 ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg
      241 ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata
      301 ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc
      361 acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg
      421 ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca
      481 aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc
      541 ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc
      601 ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt
      661 ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata
      721 tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat
      781 cggatctggt tccgcgtgga tccccggaat tcgagctcgg tacccgggga tcctctagag
      841 ctatgttacc gacccaagct ggggccgcgg cggctctggg ccggggctcg gccctggggg
      901 gcagcctgaa ccggaccccg acggggcggc cgggcggcgg cggcgggaca cgcggggcta
      961 acgggggccg ggtccccggg aatggcgcgg ggctcgggcc cggccgcctg gagcgggagg
     1021 ctgcggcagc ggcggcaacc accccggcgc ccaccgcggg ggccctctac agcggcagcg
     1081 agggcgactc ggagtcgggc gaggaggagg agctgggcgc cgagcggcgc ggcctgaagc
     1141 ggagcctgag cgagatggag atcggtatgg tggtcggtgg gcccgaggcg tcggcagcgg
     1201 ccaccggggg ctacgggccg gtgagcggcg cggtgagcgg ggccaagccg ggtaagaaga
     1261 cccggggccg cgtgaagatc aagatggagt tcatcgacaa caagctgcgg cgctacacga
     1321 ccttcagcaa gaggaagacg ggcatcatga agaaggccta tgagctgtcc acgctgacag
     1381 ggacacaggt gctgttgctg gtggccagtg agacaggcca tgtgtatacc tttgccaccc
     1441 gaaaactgca gcccatgatc accagtgaga ccggcaaggc actgattcag acctgcctca
     1501 actcgccaga ctctccaccc cgttcagacc ccacaacaga ccagagaatg agtgccactg
     1561 gctttgaaga gacagatctc acctaccagg tgtcggagtc tgacagcagt ggggagacca
     1621 aggacacact gaagccggcg ttcacagtca ccaacctgcc gggtacaacc tccaccatcc
     1681 aaacagcacc tagcacctct accaccatgc aagtcagcag cggcccctcc tttcccatca
     1741 ccaactacct ggcaccagtg tctgctagtg tcagccccag tgctgtcagc agtgccaatg
     1801 ggactgtgct gaagagtaca ggcagcggcc ctgtctcctc tgggggcctt atgcagctgc
     1861 ctaccagctt caccctcatg cctggtgggg cagtggccca gcaggtccca gtgcaggcca
     1921 ttcaagtgca ccaggcccca cagcaagcgt ctccctcccg tgacagcagc acagacctca
     1981 cgcagacctc ctccagcggg acagtgacgc tgcccgccac catcatgacg tcatccgtgc
     2041 ccacaactgt gggtggccac atgatgtacc ctagcccgca tgcggtgatg tatgccccca
     2101 cctcgggcct gggtgatggc agcctcaccg tgctgaatgc cttctcccag gcaccatcca
     2161 ccatgcaggt gtcacacagc caggtccagg agccaggtgg cgtcccccag gtgttcctga
     2221 cagcatcatc tgggacagtg cagatccctg tttcagcagt tcagctccac cagatggctg
     2281 tgatagggca gcaggccggg agcagcagca acctcaccga gctacaggtg gtgaacctgg
     2341 acaccgccca cagcaccaag agtgaatgat ccgcccgccg ccctggacag atggcccaag
     2401 ggatggcacc acttatttat tgttgccttt tcacgttttc tttacacaca cgttgactct
     2461 agagtcgact cgagcggccg catcgtgact gactgacgat ctgcctcgcg cgtttcggtg
     2521 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2581 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2641 gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga agacgaaagg
     2701 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     2761 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2821 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2881 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2941 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3001 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3061 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     3121 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     3181 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     3241 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3301 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3361 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3421 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3481 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3541 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3601 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3661 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3721 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     3781 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     3841 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     3901 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     3961 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4021 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4081 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4141 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4201 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4261 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4321 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4381 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4441 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4501 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4561 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4621 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4681 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4741 accgcataaa ttccgacacc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc
     4801 gcccggaaga gagtcaattc agggtggtga atgtgaaacc agtaacgtta tacgatgtcg
     4861 cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg
     4921 tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca
     4981 accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca
     5041 gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac
     5101 tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg
     5161 cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg
     5221 accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg
     5281 tctctgacca gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg
     5341 gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa
     5401 gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa
     5461 ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca
     5521 tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg
     5581 cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg
     5641 tagtgggata cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca
     5701 aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg
     5761 gccaggcggt gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc
     5821 tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     5881 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag
     5941 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     6001 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cggattcact
     6061 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     6121 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     6181 ttcccaacag ttgcgcagcc tgaatggcga atggcgcttt gcctggtttc cggcaccaga
     6241 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc
     6301 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat
     6361 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt
     6421 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttgg
     6481 aattagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
     6541 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc
     6601 ccgttctgga taatgt
//
