LOCUS       sequence-237155-        6025 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Foxf1 as a GST fusion protein in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6025)
  AUTHORS   Hoggatt AM, Kim JR, Ustiyan V, Ren X, Kalin TV, Kalinichenko VV, 
            Herring BP
  TITLE     The transcription factor Foxf1 binds to serum response factor and 
            myocardin to regulate gene transcription in visceral smooth muscle 
            cells.
  JOURNAL   J Biol Chem. 2013 Oct 4;288(40):28477-87. doi: 
            10.1074/jbc.M113.478974. Epub 2013 Aug 14.
  PUBMED    23946491
REFERENCE   2  (bases 1 to 6025)
  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: 
            "10.1074/jbc.M113.478974"; journalName: "J Biol Chem"; date: 
            "2013-10-4- 4"; volume: "288"; issue: "40"; pages: "28477-87"
COMMENT     Sequence Label: Foxf1 pGEX4T1
FEATURES             Location/Qualifiers
     source          1..6025
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(1..6,7..22,23..29)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    37..53
                     /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             76..729
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     687..709
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             736..753
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1893..1915)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2060..2078)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2146..2250
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2251..2319,2320..3111)
                     /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(2469..2488)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3282..3870
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3771..3790
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4024..4041
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4114..4191
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4192..5274
                     /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(4211..4230)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5323..5328,5329..5346,5347..5353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5361..5377
                     /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     5385..5401
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5385..5401
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5397..5924
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5417..5434)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5417..5433)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5426..5448)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 ttgacaatta atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca
       61 caggaaacag tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa
      121 cccactcgac ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc
      181 gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt
      241 ccttattata ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata
      301 gctgacaagc acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt
      361 gaaggagcgg ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt
      421 gaaactctca aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat
      481 cgtttatgtc ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg
      541 tatgacgctc ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa
      601 ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc
      661 agcaagtata tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat
      721 cctccaaaat cggatctggt tccgcgtgga tccccggaat tcatggaccc cgcggcggcg
      781 ggccccacca aggccaagaa gaccaacgcc ggcgtgcggc ggccggagaa gccgccctac
      841 tcgtacatcg cgctcatcgt catggctatc cagagctcgc ccagcaagcg cctgaccctc
      901 agcgagatct accagtttct tcaggcgcgc ttccccttct tccgcggcgc ctaccagggc
      961 tggaagaact ccgtgcgcca caacctgtcg ctcaacgagt gcttcatcaa actgcccaag
     1021 ggcctcgggc gacccggcaa gggccactac tggaccatcg atccggctag cgagtttatg
     1081 ttcgaggagg gctcgttccg ccggcggccg cgcggcttcc gaaggaaatg ccaggcgctc
     1141 aagcctgtgt acagcatggt gaacgggctg ggcttcaacc acctccccga cacctacggc
     1201 ttccagggct ccggaggcct ctcgtgcgcg cccaacagcc tggcgctgga gggcggcttg
     1261 ggcatgatga atggccattt ggctggcaac gtggacggca tggctttgcc cagccactcg
     1321 gtgccacacc tgccctccaa cggcggccac tcgtacatgg gtggctgcgg tggctctgcg
     1381 gccggggagt acccgcacca cgacagctcg gtgcccgctt caccgctgct gccggccggc
     1441 gccggcggag tcatggagcc gcacgccgtt tactccagct ctgcagcagc ctggccgccc
     1501 gcggcctccg cggctctcaa cagcggggcc tcctacatca agcaacagcc tctgtcccct
     1561 tgcaacccag ccgccaaccc cctgtctggc agcatctcca cgcactccct ggagcagcca
     1621 taccttcacc aaaacagtca caacgggccg gcagaactgc aaggcatccc tcggtatcac
     1681 tcacagtcgc ccagcatgtg tgacagaaag gagtttgtct tctctttcaa tgccatggcc
     1741 tcttcttcta tgcatacaac aggcggagga tcttactatc accagcaggt cacctaccaa
     1801 gacatcaagc cgtgtgtgat gtgaggtctc gagcggccgc atcgtgactg actgacgatc
     1861 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     1921 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     1981 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     2041 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     2101 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2161 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2221 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2281 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2461 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2761 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     2821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3121 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3181 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3241 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3301 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3361 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3421 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3481 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3541 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3601 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3661 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3721 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3781 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3841 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3901 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     3961 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4021 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4081 catctgtgcg gtatttcaca ccgcataaat tccgacacca tcgaatggcg caaaaccttt
     4141 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     4201 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     4261 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     4321 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     4381 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     4441 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     4501 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     4561 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     4621 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     4681 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     4741 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     4801 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     4861 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     4921 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     4981 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     5041 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     5101 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     5161 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     5221 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     5281 cgcaattaat gtaagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     5341 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     5401 ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     5461 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     5521 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg
     5581 cctggtttcc ggcaccagaa gcggtgccgg aaagctggct ggagtgcgat cttcctgagg
     5641 ccgatactgt cgtcgtcccc tcaaactggc agatgcacgg ttacgatgcg cccatctaca
     5701 ccaacgtgac ctatcccatt acggtcaatc cgccgtttgt tcccacggag aatccgacgg
     5761 gttgttactc gctcacattt aatgttgatg aaagctggct acaggaaggc cagacgcgaa
     5821 ttatttttga tggcgttgga attagcttat cgactgcacg gtgcaccaat gcttctggcg
     5881 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     5941 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     6001 ctggcaaata ttctgaaatg agctg
//
