LOCUS       sequence-205977-        8124 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 8124)
  AUTHORS   Nakamura N, Ramaswamy S, Vazquez F, Signoretti S, Loda M, Sellers WR
  TITLE     Forkhead transcription factors are critical effectors of cell death 
            and cell cycle arrest downstream of PTEN.
  JOURNAL   Mol Cell Biol. 2000 Dec . 20(23):8969-82.
  PUBMED    11073996
REFERENCE   2  (bases 1 to 8124)
  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: "Mol Cell 
            Biol. 2000 Dec . 20(23):8969-82."
COMMENT     Sequence Label: 1099 pcDNA GFP FKHR AAA
FEATURES             Location/Qualifiers
     source          1..8124
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     553..573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     647..666
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        647..665
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             685..1398
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(715..743)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1338..1358
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(3461..3479)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3462..3479)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3499..3516)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3505..3729
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3775..4203
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3862..3881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4072..4093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4212..4232)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4217..4546
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4397..4532
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4613..5407
                     /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(4667..4686)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5277..5296
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5581..5702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5618..5637)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5672..5691
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5751..5767)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5751..5767)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5764..5786)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5775..5791
                     /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(5799..5805,5806..5823,5824..5829))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5844..5865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5982..5999)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6153..6741)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6233..6252)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6912..7703,7704..7772))
                     /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     7535..7554
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7773..7877)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7952..7971)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      661 atagggagac ccaagcttgc caccatgagt aaaggagaag aactcttcac tggagttgtc
      721 ccaattcttg ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag
      781 ggtgaaggtg atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa
      841 ctacctgttc catggccaac acttgtcact actttcactt atggtgttca gtgcttttca
      901 agatacccgg atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat
      961 gtacaggaaa gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc
     1021 aagtttgaag gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa
     1081 gatggaaaca ttcttgggca caaattggaa tacaactata actcacacaa tgtatacatc
     1141 atggcagaca aacaaaagaa tggaatcaaa gctaacttca aaattagaca caacattgaa
     1201 gatggaagcg ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct
     1261 gtccttttac cagacaacca ttacctgtcc acacaatctg ccctttcgaa agatcccaac
     1321 gaaaagagag accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc
     1381 atggatgaac tatacaaaca gcagcttcag ggatccgagc tcggatctga tgtcaccatg
     1441 gccgaggcgc ctcaggtggt ggagatcgac ccggacttcg agccgctgcc ccggccgcgc
     1501 tcgtgcgcct ggccgctgcc caggccggag tttagccagt ccaactcggc cacctccagc
     1561 ccggcgccgt cgggcagcgc ggctgccaac cccgacgccg cggcgggcct gccctcggcc
     1621 tcggctgccg ctgtcagcgc cgacttcatg agcaacctga gcttgctgga ggagagcgag
     1681 gacttcccgc aggcgcccgg ctccgtggcg gcggcggtgg cggcggcggc cgccgcggcc
     1741 gccaccgggg ggctgtgcgg ggacttccag ggcccggagg cgggctgcct gcacccagcg
     1801 ccaccgcagc ccccgccgcc cgggccgctg tcgcagcacc cgccggtgcc ccccgccgcc
     1861 gctgggccgc tcgcggggca gccgcgcaag agcagctcgt cccgccgcaa cgcgtggggc
     1921 aacctgtcct acgccgacct catcaccaag gccatcgaga gctcggcgga gaagcggctc
     1981 acgctgtcgc agatctacga gtggatggtc aagagcgtgc cctacttcaa ggataagggt
     2041 gacagcaaca gctcggcggg ctggaagaat tcaattcgtc ataatctgtc cctacacagc
     2101 aagttcattc gtgtgcagaa tgaaggaact ggaaaaagtt cttggtggat gctcaatcca
     2161 gagggtggca agagcgggaa atctcctagg agaagagctg cagccatgga caacaacagt
     2221 aaatttgcta agagccgaag ccgagctgcc aagaagaaag catctctcca gtctggccag
     2281 gagggtgctg gggacagccc tggatcacag ttttccaaat ggcctgcaag ccctggctct
     2341 cacagcaatg atgactttga taactggagt acatttcgcc ctcgaactag cgcaaatgct
     2401 agtactatta gtgggagact ctcacccatt atgaccgaac aggatgatct tggagaaggg
     2461 gatgtgcatt ctatggtgta cccgccatct gccgcaaaga tggcctctac tttacccagt
     2521 ctgtctgaga taagcaatcc cgaaaacatg gaaaatcttt tggataatct caaccttctc
     2581 tcatcaccaa catcattaac tgtttcgacc cagtcctcac ctggcaccat gatgcagcag
     2641 acgccgtgct actcgtttgc gccaccaaac accagtttga attcacccag cccaaactac
     2701 caaaaatata catatggcca atccagcatg agccctttgc cccagatgcc tatacaaaca
     2761 cttcaggaca ataagtcgag ttatggaggt atgagtcagt ataactgtgc gcctggactc
     2821 ttgaaggagt tgctgacttc tgactctcct ccccataatg acattatgac accagttgat
     2881 cctggggtag cccagcccaa cagccgggtt ctgggccaga acgtcatgat gggccctaat
     2941 tcggtcatgt caacctatgg cagccaggca tctcataaca aaatgatgaa tcccagctcc
     3001 catacccacc ctggacatgc tcagcagaca tctgcagtca acgggcgtcc cctgccccac
     3061 acggtaagca ccatgcccca cacctcgggt atgaaccgcc tgacccaagt gaagacacct
     3121 gtacaagtgc ctctgcccca ccccatgcag atgagtgccc tggggggcta ctcctccgtg
     3181 agcagctgca atggctatgg cagaatgggc cttctccacc aggagaagct cccaagtgac
     3241 ttggatggca tgttcattga gcgcttagac tgtgacatgg aatccatcat tcggaatgac
     3301 ctcatggatg gagatacatt ggattttaac tttgacaatg tgttgcccaa ccaaagcttc
     3361 ccacacagtg tcaagacaac gacacatagc tgggtgtcag gctgagggtt agtgagcagg
     3421 ttacacttaa aagttcgagc atgcatctag agggccctat tctatagtgt cacctaaatg
     3481 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     3541 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     3601 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     3661 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     3721 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3781 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3841 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3901 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3961 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4021 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4081 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4141 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4201 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4261 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4321 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4381 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4441 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     4501 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4561 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4621 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4681 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4741 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4801 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4861 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4921 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4981 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     5041 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     5101 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     5161 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     5221 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     5281 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5341 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5401 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5461 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5521 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5581 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5641 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5701 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5761 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5821 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5881 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5941 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     6001 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     6061 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     6121 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     6181 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6241 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6301 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6361 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6421 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6481 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6541 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6601 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6661 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6721 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6781 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6841 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6901 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6961 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     7021 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     7081 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     7141 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     7201 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     7261 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     7321 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     7381 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     7441 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     7501 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     7561 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     7621 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     7681 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     7741 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7801 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7861 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     7921 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7981 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt
     8041 aagctacaac aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc
     8101 gttttgcgct gcttcgcgat gtac
//
