LOCUS       sequence-320217-        6656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ETS gene FLI1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6656)
  AUTHORS   Selvaraj N, Kedage V, Hollenhorst PC
  TITLE     Comparison of MAPK specificity across the ETS transcription factor 
            family identifies a high-affinity ERK interaction required for ERG 
            function in prostate cells.
  JOURNAL   Cell Commun Signal. 2015 Feb 19;13:12. doi: 
            10.1186/s12964-015-0089-7.
  PUBMED    25885538
REFERENCE   2  (bases 1 to 6656)
  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: "Cell Commun
            Signal."; date: "2015-02-19"; volume: "13:12. doi"; pages: " 
            10.1186/s12964-015-0089-7"
COMMENT     Sequence Label: FLI1_pet28a
FEATURES             Location/Qualifiers
     source          1..6656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1534..1551)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(1561..1578)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1597..1619
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1634..1658
                     /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     complement(1658..1677)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1659..1677)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1761..1780)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1986..2063
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2064..3146
                     /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(2083..2102)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3955..4146
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4140..4162)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4248..4387
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4303..4322
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4402..4419)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4573..5161)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4653..4672)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5272..5291)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5283..6098
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5283..6098
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5356..5375)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6190..6645)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6327..6348)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6539..6558
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gctagtagta
      181 gctgcctaag tgtgaaggca cgtgggtgtt aggatggcgg gggacgttgg ggttggggta
      241 gattcccccc gtgggggagg tccagtattg tgatgcggct ccaaagaagc tggaggaagt
      301 gacaggcatg gaggatggat ggggagggac aaagttcacc ttctgctggt gggcatggta
      361 ggaaggcatg taggagatgt cagaagggta cttgtacatg gacgactcgg tcggatgtgg
      421 ctgcagagcc tgggcaatgc cgtggaagtc aaatttgtaa gcatatcttt tgccgtgcac
      481 tttggtcata atgtttttat catagtaata acggagggcc cggctcagct tgtcgtaatt
      541 catgttgggc ttgcttttcc gctcgcccca gcgcctggcc acctcatcgg ggtccgtcat
      601 tttgaactcc ccgttggtcc cctcccaggt gatacagctg gcgttggcgc tgtcggagag
      661 cagctccagg aggaattgcc acagctggat ctgcccgctt ccagggttgg ctaggcgact
      721 gctggtcggg cccaggatct gatacggatc tggctggggc cgttgctctg tattcttact
      781 gatcgtttgt gcccctccaa ggggaggact tttgttgagg ccagaattca tgttattgcc
      841 ccaagctcct cttctgactg agtcataaga agggtcttct ttgacactca atcgtgagga
      901 ttggtcggtg tgggaggttg tattataggc cagcagtgaa ctttccctga ggtaactgag
      961 gtgtgacaac agcacttccg tgttgtagag ggtggtggcg cggaggaagt cctccttgtt
     1021 cattttacac agttccttgc catccatgtt ctggaaaaag gatgtgtcga tctccatcaa
     1081 gctgtactcc tttatggccc actccagcca ttgcctcaca tgctcctgtg tccacagtgt
     1141 ggggtctgcg gggacgatga ctctcctctc gttggtggtc atgttgggag gaggggggcc
     1201 attcttctcg tccatatagc tgttgtagtt catggggttg gactcgcctc cgcccaccag
     1261 cttgctgcat ttgctaacgc tgcagtccac cggagactcc ctggatccat tcatgtggtc
     1321 atactcccgc ttgacgttga ccctcactgg ctgattgatc cactcctgct gtggtgggag
     1381 ggggttgatc ttgtggggct gcccgtagtc aggactcccc gaggcagtca tgtcggcctt
     1441 ggggagatgg gctgccgctc cgtacgctga gtcaaagagg gactggtcgt cgctcaccac
     1501 cgacagagcc tccttaatag tcccgtccat atggctgccg cgcggcacca ggccgctgct
     1561 gtgatgatga tgatgatggc tgctgcccat ggtatatctc cttcttaaag ttaaacaaaa
     1621 ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt
     1681 tcgcgggatc gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc
     1741 cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag atcgggctcg
     1801 ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc ccgtggccgg
     1861 gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg tgctcaacgg
     1921 cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag agcgtcgaga
     1981 tcccggacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     2041 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     2101 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     2161 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     2221 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     2281 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     2341 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     2401 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     2461 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     2521 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     2581 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     2641 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     2701 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     2761 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     2821 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     2881 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     2941 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     3001 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     3061 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     3121 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     3181 taggcaccgg gatctcgacc gatgcccttg agagccttca acccagtcag ctccttccgg
     3241 tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc
     3301 gtaggacagg tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc
     3361 gcgacgatga tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc
     3421 ttcgtcactg gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg
     3481 gcggccccac gggtgcgcat gatcgtgctc ctgtcgttga ggacccggct aggctggcgg
     3541 ggttgcctta ctggttagca gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc
     3601 tgctgcaaaa cgtctgcgac ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg
     3661 taaagtctgg aaacgcggaa gtcagcgccc tgcaccatta tgttccggat ctgcatcgca
     3721 ggatgctgct ggctaccctg tggaacacct acatctgtat taacgaagcg ctggcattga
     3781 ccctgagtga tttttctctg gtcccgccgc atccataccg ccagttgttt accctcacaa
     3841 cgttccagta accgggcatg ttcatcatca gtaacccgta tcgtgagcat cctctctcgt
     3901 ttcatcggta tcattacccc catgaacaga aatccccctt acacggaggc atcagtgacc
     3961 aaacaggaaa aaaccgccct taacatggcc cgctttatca gaagccagac attaacgctt
     4021 ctggagaaac tcaacgagct ggacgcggat gaacaggcag acatctgtga atcgcttcac
     4081 gaccacgctg atgagcttta ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     4141 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4201 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     4261 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     4321 tactgagagt gcaccattgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4381 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4441 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4501 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4561 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4621 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4681 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4741 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4801 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4861 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4921 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4981 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5041 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5101 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5161 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5221 agggattttg gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     5281 ttatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg
     5341 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     5401 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     5461 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     5521 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     5581 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     5641 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     5701 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     5761 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     5821 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     5881 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     5941 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     6001 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     6061 aattgcagtt tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac
     6121 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6181 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     6241 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     6301 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     6361 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     6421 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     6481 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     6541 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     6601 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//
