LOCUS       sequence-131703-        7423 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 7423)
  AUTHORS   Tang ED, Nunez G, Barr FG, Guan KL
  TITLE     Negative regulation of the forkhead transcription factor FKHR by 
            Akt.
  JOURNAL   J Biol Chem. 1999 Jun 11. 274(24):16741-6.
  PUBMED    10358014
REFERENCE   2  (bases 1 to 7423)
  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: "J Biol 
            Chem. 1999 Jun 11. 274(24):16741-6."
COMMENT     Sequence Label: pcDNA3 Flag FKHR
FEATURES             Location/Qualifiers
     source          1..7423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             901..924
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2975..2993)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2976..2993)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3013..3030)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3019..3243
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3289..3717
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3376..3395)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3586..3607
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3726..3746)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3731..4061
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3912..4047
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     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 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttaccatg
      901 gactacaagg acgacgatga caagggatcc gagctcggat ctgatgtcac catggccgag
      961 gcgcctcagg tggtggagat cgacccggac ttcgagccgc tgccccggcc gcgctcgtgc
     1021 acctggccgc tgcccaggcc ggagtttagc cagtccaact cggccacctc cagcccggcg
     1081 ccgtcgggca gcgcggctgc caaccccgac gccgcggcgg gcctgccctc ggcctcggct
     1141 gccgctgtca gcgccgactt catgagcaac ctgagcttgc tggaggagag cgaggacttc
     1201 ccgcaggcgc ccggctccgt ggcggcggcg gtggcggcgg cggccgccgc ggccgccacc
     1261 ggggggctgt gcggggactt ccagggcccg gaggcgggct gcctgcaccc agcgccaccg
     1321 cagcccccgc cgcccgggcc gctgtcgcag cacccgccgg tgccccccgc cgccgctggg
     1381 ccgctcgcgg ggcagccgcg caagagcagc tcgtcccgcc gcaacgcgtg gggcaacctg
     1441 tcctacgccg acctcatcac caaggccatc gagagctcgg cggagaagcg gctcacgctg
     1501 tcgcagatct acgagtggat ggtcaagagc gtgccctact tcaaggataa gggtgacagc
     1561 aacagctcgg cgggctggaa gaattcaatt cgtcataatc tgtccctaca cagcaagttc
     1621 attcgtgtgc agaatgaagg aactggaaaa agttcttggt ggatgctcaa tccagagggt
     1681 ggcaagagcg ggaaatctcc taggagaaga gctgcatcca tggacaacaa cagtaaattt
     1741 gctaagagcc gaagccgagc tgccaagaag aaagcatctc tccagtctgg ccaggagggt
     1801 gctggggaca gccctggatc acagttttcc aaatggcctg caagccctgg ctctcacagc
     1861 aatgatgact ttgataactg gagtacattt cgccctcgaa ctagctcaaa tgctagtact
     1921 attagtggga gactctcacc cattatgacc gaacaggatg atcttggaga aggggatgtg
     1981 cattctatgg tgtacccgcc atctgccgca aagatggcct ctactttacc cagtctgtct
     2041 gagataagca atcccgaaaa catggaaaat cttttggata atctcaacct tctctcatca
     2101 ccaacatcat taactgtttc gacccagtcc tcacctggca ccatgatgca gcagacgccg
     2161 tgctactcgt ttgcgccacc aaacaccagt ttgaattcac ccagcccaaa ctaccaaaaa
     2221 tatacatatg gccaatccag catgagccct ttgccccaga tgcctataca aacacttcag
     2281 gacaataagt cgagttatgg aggtatgagt cagtataact gtgcgcctgg actcttgaag
     2341 gagttgctga cttctgactc tcctccccat aatgacatta tgacaccagt tgatcctggg
     2401 gtagcccagc ccaacagccg ggttctgggc cagaacgtca tgatgggccc taattcggtc
     2461 atgtcaacct atggcagcca ggcatctcat aacaaaatga tgaatcccag ctcccatacc
     2521 caccctggac atgctcagca gacatctgca gtcaacgggc gtcccctgcc ccacacggta
     2581 agcaccatgc cccacacctc gggtatgaac cgcctgaccc aagtgaagac acctgtacaa
     2641 gtgcctctgc cccaccccat gcagatgagt gccctggggg gctactcctc cgtgagcagc
     2701 tgcaatggct atggcagaat gggccttctc caccaggaga agctcccaag tgacttggat
     2761 ggcatgttca ttgagcgctt agactgtgac atggaatcca tcattcggaa tgacctcatg
     2821 gatggagata cattggattt taactttgac aatgtgttgc ccaaccaaag cttcccacac
     2881 agtgtcaaga caacgacaca tagctgggtg tcaggctgag ggttagtgag caggttacac
     2941 ttaaaagttc gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag
     3001 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     3061 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     3121 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     3181 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     3241 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     3301 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3361 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3421 ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc
     3481 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3541 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3601 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg
     3661 ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     3721 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 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 caatgctcac 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 actagaagga 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 gtc
//
