LOCUS       Exported                8124 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    1098 pcDNA GFP FKHR
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 May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8124
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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..1616
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(933..961)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1556..1576
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(3679..3697)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3680..3697)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3717..3734)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3723..3947
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3993..4421
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4080..4099)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4290..4311
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4430..4450)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4435..4764
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4615..4750
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4677..4696
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4831..5625
                     /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(4885..4904)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5495..5514
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5799..5920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5836..5855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5890..5909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5969..5985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5969..5985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5982..6004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5993..6009
                     /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(6017..6047)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6062..6083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6200..6217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6371..6959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6451..6470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7130..7990)
                     /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     7753..7772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7991..8095)
                     /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 aagcttgcca
      901 ccatgagtaa aggagaagaa ctcttcactg gagttgtccc aattcttgtt gaattagatg
      961 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
     1021 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac
     1081 ttgtcactac tttcacttat ggtgttcagt gcttttcaag atacccggat catatgaaac
     1141 ggcatgactt tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt
     1201 tcaaagatga cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg
     1261 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttgggcaca
     1321 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
     1381 gaatcaaagc taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag
     1441 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     1501 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc
     1561 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaacagc
     1621 agcttcaggg atccgagctc ggatctgatg tcaccatggc cgaggcgcct caggtggtgg
     1681 agatcgaccc ggacttcgag ccgctgcccc ggccgcgctc gtgcacctgg ccgctgccca
     1741 ggccggagtt tagccagtcc aactcggcca cctccagccc ggcgccgtcg ggcagcgcgg
     1801 ctgccaaccc cgacgccgcg gcgggcctgc cctcggcctc ggctgccgct gtcagcgccg
     1861 acttcatgag caacctgagc ttgctggagg agagcgagga cttcccgcag gcgcccggct
     1921 ccgtggcggc ggcggtggcg gcggcggccg ccgcggccgc caccgggggg ctgtgcgggg
     1981 acttccaggg cccggaggcg ggctgcctgc acccagcgcc accgcagccc ccgccgcccg
     2041 ggccgctgtc gcagcacccg ccggtgcccc ccgccgccgc tgggccgctc gcggggcagc
     2101 cgcgcaagag cagctcgtcc cgccgcaacg cgtggggcaa cctgtcctac gccgacctca
     2161 tcaccaaggc catcgagagc tcggcggaga agcggctcac gctgtcgcag atctacgagt
     2221 ggatggtcaa gagcgtgccc tacttcaagg ataagggtga cagcaacagc tcggcgggct
     2281 ggaagaattc aattcgtcat aatctgtccc tacacagcaa gttcattcgt gtgcagaatg
     2341 aaggaactgg aaaaagttct tggtggatgc tcaatccaga gggtggcaag agcgggaaat
     2401 ctcctaggag aagagctgca tccatggaca acaacagtaa atttgctaag agccgaagcc
     2461 gagctgccaa gaagaaagca tctctccagt ctggccagga gggtgctggg gacagccctg
     2521 gatcacagtt ttccaaatgg cctgcaagcc ctggctctca cagcaatgat gactttgata
     2581 actggagtac atttcgccct cgaactagct caaatgctag tactattagt gggagactct
     2641 cacccattat gaccgaacag gatgatcttg gagaagggga tgtgcattct atggtgtacc
     2701 cgccatctgc cgcaaagatg gcctctactt tacccagtct gtctgagata agcaatcccg
     2761 aaaacatgga aaatcttttg gataatctca accttctctc atcaccaaca tcattaactg
     2821 tttcgaccca gtcctcacct ggcaccatga tgcagcagac gccgtgctac tcgtttgcgc
     2881 caccaaacac cagtttgaat tcacccagcc caaactacca aaaatataca tatggccaat
     2941 ccagcatgag ccctttgccc cagatgccta tacaaacact tcaggacaat aagtcgagtt
     3001 atggaggtat gagtcagtat aactgtgcgc ctggactctt gaaggagttg ctgacttctg
     3061 actctcctcc ccataatgac attatgacac cagttgatcc tggggtagcc cagcccaaca
     3121 gccgggttct gggccagaac gtcatgatgg gccctaattc ggtcatgtca acctatggca
     3181 gccaggcatc tcataacaaa atgatgaatc ccagctccca tacccaccct ggacatgctc
     3241 agcagacatc tgcagtcaac gggcgtcccc tgccccacac ggtaagcacc atgccccaca
     3301 cctcgggtat gaaccgcctg acccaagtga agacacctgt acaagtgcct ctgccccacc
     3361 ccatgcagat gagtgccctg gggggctact cctccgtgag cagctgcaat ggctatggca
     3421 gaatgggcct tctccaccag gagaagctcc caagtgactt ggatggcatg ttcattgagc
     3481 gcttagactg tgacatggaa tccatcattc ggaatgacct catggatgga gatacattgg
     3541 attttaactt tgacaatgtg ttgcccaacc aaagcttccc acacagtgtc aagacaacga
     3601 cacatagctg ggtgtcaggc tgagggttag tgagcaggtt acacttaaaa gttcgagcat
     3661 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     3721 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3781 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3841 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3901 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3961 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     4021 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4081 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4141 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4201 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4261 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4321 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4381 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4441 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4501 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4561 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4621 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4681 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4741 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4801 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     4861 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     4921 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     4981 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     5041 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     5101 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     5161 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     5221 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     5281 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     5341 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     5401 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     5461 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     5521 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     5581 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     5641 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     5701 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     5761 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     5821 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     5881 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     5941 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6001 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6061 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6121 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6181 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6241 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6301 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6361 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6421 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6481 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6541 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6601 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6661 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6721 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6781 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6841 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6901 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6961 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7021 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7081 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7141 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7201 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7261 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7321 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7381 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7441 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7501 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7561 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7621 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7681 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7741 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7801 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7861 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7921 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7981 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8041 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8101 ttccccgaaa agtgccacct gacg
//
