LOCUS       sequence-137167-        7422 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 7422)
  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 7422)
  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 AAA mutant
FEATURES             Location/Qualifiers
     source          1..7422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..59
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        40..58
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             78..101
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2152..2170)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2153..2170)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2190..2207)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2196..2420
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2466..2894
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2553..2572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2763..2784
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2903..2923)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2908..3237
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3088..3223
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3304..4098
                     /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(3358..3377)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3968..3987
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4272..4393
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4309..4328)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4363..4382
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4442..4458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4442..4458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4455..4477)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4466..4482
                     /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(4490..4496,4497..4514,4515..4520))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4535..4556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4673..4690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4844..5432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4924..4943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5603..6394,6395..6463))
                     /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     6226..6245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6464..6568)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6643..6662)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6834..7213
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7214..7417
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7368..7388
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
       61 gacccaagct taccatggac tacaaggacg acgatgacaa gggatccgag ctcggatctg
      121 atgtcaccat ggccgaggcg cctcaggtgg tggagatcga cccggacttc gagccgctgc
      181 cccggccgcg ctcgtgcgcc tggccgctgc ccaggccgga gtttagccag tccaactcgg
      241 ccacctccag cccggcgccg tcgggcagcg cggctgccaa ccccgacgcc gcggcgggcc
      301 tgccctcggc ctcggctgcc gctgtcagcg ccgacttcat gagcaacctg agcttgctgg
      361 aggagagcga ggacttcccg caggcgcccg gctccgtggc ggcggcggtg gcggcggcgg
      421 ccgccgcggc cgccaccggg gggctgtgcg gggacttcca gggcccggag gcgggctgcc
      481 tgcacccagc gccaccgcag cccccgccgc ccgggccgct gtcgcagcac ccgccggtgc
      541 cccccgccgc cgctgggccg ctcgcggggc agccgcgcaa gagcagctcg tcccgccgca
      601 acgcgtgggg caacctgtcc tacgccgacc tcatcaccaa ggccatcgag agctcggcgg
      661 agaagcggct cacgctgtcg cagatctacg agtggatggt caagagcgtg ccctacttca
      721 aggataaggg tgacagcaac agctcggcgg gctggaagaa ttcaattcgt cataatctgt
      781 ccctacacag caagttcatt cgtgtgcaga atgaaggaac tggaaaaagt tcttggtgga
      841 tgctcaatcc agagggtggc aagagcggga aatctcctag gagaagagct gcagccatgg
      901 acaacaacag taaatttgct aagagccgaa gccgagctgc caagaagaaa gcatctctcc
      961 agtctggcca ggagggtgct ggggacagcc ctggatcaca gttttccaaa tggcctgcaa
     1021 gccctggctc tcacagcaat gatgactttg ataactggag tacatttcgc cctcgaacta
     1081 gcgcaaatgc tagtactatt agtgggagac tctcacccat tatgaccgaa caggatgatc
     1141 ttggagaagg ggatgtgcat tctatggtgt acccgccatc tgccgcaaag atggcctcta
     1201 ctttacccag tctgtctgag ataagcaatc ccgaaaacat ggaaaatctt ttggataatc
     1261 tcaaccttct ctcatcacca acatcattaa ctgtttcgac ccagtcctca cctggcacca
     1321 tgatgcagca gacgccgtgc tactcgtttg cgccaccaaa caccagtttg aattcaccca
     1381 gcccaaacta ccaaaaatat acatatggcc aatccagcat gagccctttg ccccagatgc
     1441 ctatacaaac acttcaggac aataagtcga gttatggagg tatgagtcag tataactgtg
     1501 cgcctggact cttgaaggag ttgctgactt ctgactctcc tccccataat gacattatga
     1561 caccagttga tcctggggta gcccagccca acagccgggt tctgggccag aacgtcatga
     1621 tgggccctaa ttcggtcatg tcaacctatg gcagccaggc atctcataac aaaatgatga
     1681 atcccagctc ccatacccac cctggacatg ctcagcagac atctgcagtc aacgggcgtc
     1741 ccctgcccca cacggtaagc accatgcccc acacctcggg tatgaaccgc ctgacccaag
     1801 tgaagacacc tgtacaagtg cctctgcccc accccatgca gatgagtgcc ctggggggct
     1861 actcctccgt gagcagctgc aatggctatg gcagaatggg ccttctccac caggagaagc
     1921 tcccaagtga cttggatggc atgttcattg agcgcttaga ctgtgacatg gaatccatca
     1981 ttcggaatga cctcatggat ggagatacat tggattttaa ctttgacaat gtgttgccca
     2041 accaaagctt cccacacagt gtcaagacaa cgacacatag ctgggtgtca ggctgagggt
     2101 tagtgagcag gttacactta aaagttcgag catgcatcta gagggcccta ttctatagtg
     2161 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2221 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2281 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2341 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2401 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     2461 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2521 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2581 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2641 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2701 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2761 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2821 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2881 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2941 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3001 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3061 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3121 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     3181 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3241 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     3301 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3361 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3421 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3481 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3541 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3601 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3661 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3721 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3781 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     3841 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3901 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     3961 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4021 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4081 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4141 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4201 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4261 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4321 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4381 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4441 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4501 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4561 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4621 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4681 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4741 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4801 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4861 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4921 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4981 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5041 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5101 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5161 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5221 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5281 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5341 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5401 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5461 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     5521 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     5581 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     5641 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5701 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5761 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5821 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5881 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5941 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     6001 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     6061 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     6121 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     6181 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     6241 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     6301 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     6361 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     6421 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     6481 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6541 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
     6601 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
     6661 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
     6721 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
     6781 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
     6841 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     6901 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     6961 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     7021 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     7081 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     7141 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
     7201 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
     7261 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     7321 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     7381 taggcgtgta cggtgggagg tctatataag cagagctctc tg
//
