LOCUS       Exported                7654 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    AIRAP 11: PSMD14.pEYFP.V1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7654)
  AUTHORS   Stanhill A, Haynes CM, Zhang Y, Min G, Steele MC, Kalinina J, 
            Martinez E, Pickart CM, Kong XP, Ron D
  TITLE     An arsenite-inducible 19S regulatory particle-associated protein 
            adapts proteasomes to proteotoxicity.
  JOURNAL   Mol Cell. 2006 Sep 15. 23(6):875-85.
  PUBMED    16973439
REFERENCE   2  (bases 1 to 7654)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7654
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        24..1205
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          254..1196
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1153..1173
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    2177..2229
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2237..2246
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2243..2962
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2288..2309)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2549..2568)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2896..2917
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(2998..3016)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2999..3016)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3036..3053)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3042..3266
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3312..3740
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3399..3418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3609..3630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3749..3769)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3754..4084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3935..4070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3997..4016
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4151..4945
                     /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(4205..4224)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4815..4834
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5119..5240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5156..5175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5210..5229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5289..5305)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5289..5305)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5302..5324)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5313..5329
                     /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(5337..5367)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5382..5403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5520..5537)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5691..6279)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5771..5790)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6450..7310)
                     /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     7073..7092
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7311..7415)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7490..7509)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aggtggagaa gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc
       61 cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg
      121 gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg
      181 gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc
      241 cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta
      301 tggcccttgc gtgccttgaa ttacttccac gcccctggct gcagtacgtg attcttgatc
      361 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
      421 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
      481 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
      541 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
      601 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
      661 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
      721 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
      781 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
      841 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
      901 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
      961 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     1021 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     1081 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     1141 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     1201 cgtgaggaat tggcgaagct tcgaattcca gttactgaac agaaatatgg acagacttct
     1261 tagacttgga ggaggtatgc ctggactggg ccagggccca cctacagatg ctcctgccgt
     1321 ggacacagca gaacaagttt atatctcttc cttggccttg ctaaagatgt taaaacatgg
     1381 tcgtgctgga gtgcctatgg aagttatggg tctaatgctt ggtgaatttg ttgatgatta
     1441 caccgtcaga gtgattgatg tgtttgctat gccacagtca ggaactggtg tcagtgttga
     1501 agcagttgat ccagtgttcc aagccaaaat gttggatatg ctgaaacaaa caggaaggcc
     1561 cgagatggtt gttggttggt atcacagtca ccctggcttt ggctgttggc tttctggtgt
     1621 ggatatcaac actcagcaga gctttgaagc cttgtcggag agagctgtgg cagtggttgt
     1681 ggatcccatt cagagtgtaa aaggaaaggt tgttattgat gccttcagac tgatcaatgc
     1741 taatatgatg gtcttaggac atgaaccaag acaaacgact tcaaatctgg gccacttaaa
     1801 caagccatct atccaggcat taattcacgg actaaacaga cattattact ccatcactat
     1861 taattaccgg aaaaatgaac tggaacagaa gatgctgtta aatttgcata agaagagttg
     1921 gatggaagga ttgacacttc aggactacag tgaacactgt aaacacaatg aatcggtggt
     1981 aaaagaaatg ttggaattag ccaagaatta taataaggct gtggaagaag aagataagat
     2041 gacacctgaa cagctggcaa taaagaatgt tggcaagcag gatcccaaac gtcatttgga
     2101 agaacatgtg gatgtgctta tgacttcaaa tattgtccag tgtttggccg caatgttgga
     2161 tactgttgta tttaaactcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
     2221 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     2281 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2341 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2401 gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc
     2461 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2521 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2581 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2641 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2701 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2761 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2821 cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa
     2881 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2941 catggacgag ctgtacaagt aaagcggccg ctcgagcatg catctagagg gccctattct
     3001 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     3061 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3121 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3181 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3241 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3301 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3361 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3421 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3481 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3541 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3601 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3661 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     3721 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3781 ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3901 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3961 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4021 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4081 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4141 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4201 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     4261 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     4321 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     4381 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     4441 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     4501 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     4561 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     4621 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     4681 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     4741 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     4801 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     4861 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     4921 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     4981 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5041 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5101 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5161 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5221 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5281 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5341 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5521 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5581 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5641 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5881 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6121 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6301 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6361 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     6421 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6481 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6541 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6601 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6661 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6721 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6781 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6841 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6901 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6961 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7021 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7081 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7141 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7201 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7261 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7321 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     7381 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     7441 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     7501 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     7561 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7621 atctgcttag ggttaggcgt tttgcgctgc ttcg
//
