LOCUS       sequence-222771-        7421 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 7421)
  AUTHORS   Ramaswamy S, Nakamura N, Sansal I, Bergeron L, Sellers WR
  TITLE     A novel mechanism of gene regulation and tumor suppression by the 
            transcription factor FKHR.
  JOURNAL   Cancer Cell. 2002 Jul . 2(1):81-91.
  PUBMED    12150827
REFERENCE   2  (bases 1 to 7421)
  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: "Cancer 
            Cell. 2002 Jul . 2(1):81-91."
COMMENT     Sequence Label: 1319 pcDNA3 flag FKHRL1 AAA
FEATURES             Location/Qualifiers
     source          1..7421
                     /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             903..926
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2976..2994)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2977..2994)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3014..3031)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3020..3244
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3290..3718
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3377..3396)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3587..3608
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3727..3747)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3732..4061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3912..4047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3974..3993
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4128..4922
                     /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(4182..4201)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4792..4811
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5096..5217
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5133..5152)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5187..5206
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5266..5282)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5266..5282)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5279..5301)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5290..5306
                     /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(5314..5320,5321..5338,5339..5344))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5359..5380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5497..5514)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5668..6256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5748..5767)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6427..7218,7219..7287))
                     /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     7050..7069
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7288..7392)
                     /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 aagcttacca
      901 tggactacaa ggacgacgat gacaagggat ccatggcaga ggcaccggct tccccggccc
      961 cgctctctcc gctcgaagtg gagctggacc cggagttcga gccccagagc cgtccgcgat
     1021 cctgtgcgtg gcccctgcaa aggccggagc tccaagcgag ccctgccaag ccctcggggg
     1081 agacggccgc tgactccatg atccccgagg aggaggacga tgaagacgac gaggacggcg
     1141 ggggacgggc cggctcggcc atggcgatcg gcggcggcgg cgggagcggc acgctgggct
     1201 ccgggctgct ccttgaggac tcggcccggg tgctggcacc cggagggcaa gaccccgggt
     1261 ctgggccagc caccgcggcg ggcgggctga gcgggggtac acaggcgctg ctgcagcctc
     1321 agcaaccgct gccaccgccg cagccggggg cggctggggg ctccgggcag ccgaggaaat
     1381 gttcgtcgcg gcggaacgcc tggggaaacc tgtcctacgc ggacctgatc acccgcgcca
     1441 tcgagagctc cccggacaaa cggctcactc tgtcccagat ctacgagtgg atggtgcgtt
     1501 gcgtgcccta cttcaaggat aagggcgaca gcaacagctc tgccggctgg aagaactcca
     1561 tccggcacaa cctgtcactg catagtcgat tcatgcgggt ccagaatgag ggaactggca
     1621 agagctcttg gtggatcatc aaccctgatg gggggaagag cggaaaagcc ccccggcggc
     1681 gggctgtcgc aatggacaat agcaacaagt ataccaagag ccgtggccgc gcagccaaga
     1741 agaaggcagc cctgcagaca gcccccgaat cagctgacga cagtccctcc cagctctcca
     1801 agtggcctgg cagccccacg tcacgcagca gtgatgagct ggatgcgtgg acggacttcc
     1861 gttcacgcac caatgctaac gccagcacag tcagtggccg cctgtcgccc atcatggcaa
     1921 gcacagagtt ggatgaagtc caggacgatg atgcgcctct ctcgcccatg ctctacagca
     1981 gctcagccag cctgtcacct tcagtaagca agccgtgcac ggtggaactg ccacggctga
     2041 ctgatatggc aggcaccatg aatctgaatg atgggctgac tgaaaacctc atggacgacc
     2101 tgctggataa catcacgctc ccgccatccc agccatcgcc cactggggga ctcatgcagc
     2161 ggagctctag cttcccgtat accaccaagg gctcgggcct gggctcccca accagctcct
     2221 ttaacagcac ggtgttcgga ccttcatctc tgaactccct acgccagtct cccatgcaga
     2281 ccatccaaga gaacaagcca gctaccttct cttccatgtc acactatggt aaccagacac
     2341 tccaggacct gctcacttcg gactcactta gccacagcga tgtcatgatg acacagtcgg
     2401 accccttgat gtctcaggcc agcaccgctg tgtctgccca gaattcccgc cggaacgtga
     2461 tgcttcgcaa tgatccgatg atgtcctttg ctgcccagcc taaccaggga agtttggtca
     2521 atcagaactt gctccaccac cagcaccaaa cccagggcgc tcttggtggc agccgtgcct
     2581 tgtcgaattc tgtcagcaac atgggcttga gtgagtccag cagccttggg tcagccaaac
     2641 accagcagca gtctcctgtc agccagtcta tgcaaaccct ctcggactct ctctcaggct
     2701 cctccttgta ctcaactagt gcaaacctgc ccgtcatggg ccatgagaag ttccccagcg
     2761 acttggacct ggacatgttc aatgggagct tggaatgtga catggagtcc attatccgta
     2821 gtgaactcat ggatgctgat gggttggatt ttaactttga ttccctcatc tccacacaga
     2881 atgttgttgg tttgaacgtg gggaacttca ctggtgctaa gcaggcctca tctcagagct
     2941 gggtgccagg ctgactcgag tctagagggc cctattctat agtgtcacct aaatgctaga
     3001 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     3061 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     3121 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     3181 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     3241 atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt
     3301 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3361 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3421 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3481 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3541 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3601 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3661 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
     3721 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3781 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3841 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3901 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3961 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
     4021 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
     4081 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4141 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4201 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4261 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
     4321 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4381 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     4441 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     4501 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     4561 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     4621 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
     4681 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     4741 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     4801 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     4861 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     4921 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     4981 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5041 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     5101 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5161 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5221 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     5281 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5341 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5401 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     5461 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     5521 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     5581 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5641 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5701 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5761 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5821 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5881 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5941 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6001 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6061 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6121 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6181 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6241 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6301 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6361 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6421 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6481 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6541 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6601 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6661 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6721 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6781 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6841 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6901 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6961 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7021 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7081 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7141 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7201 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7261 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7321 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7381 aggggttccg cgcacatttc cccgaaaagt gccacctgac g
//
