LOCUS       Exported                5562 bp ds-DNA     circular SYN 14-FEB-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5562)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Feb 21, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     pHW393 pPD117.01-Prab-3-Gal4(sk)-VP64
            use (2.2) clone
            clone (2.1), sequenced with oHW10f, 11r and oHW12f (laragen #105015,
            at 20150722) ,  sequence sof  the Prab-3-Gal4(sk)-VP64 section is 
            correct, and the Decoy seqeuence and let-858 sequence from pPD117.01
            is confirmed. But the result of oHW12f sequence revealed that (2.1) 
            is missing a single ""G"" right after the stop codon ""TAA"" and 
            thus destoy the KpnI site  (further confirmedy with oHW233r laragen 
            #105284 at 2015072)
            
            Clone (2.2) (the Gal4-(sk)-VP64 section) is confirmed by onfirmedy 
            with oHW233r (in the let-858 3'UTR)  laragen #105284 at 20150727) 
            and sequence with oHW10f at 20150728, #105308)
            Insert: ~2.0 kb of Prab-3-Gal4(sk)-VP64, PCR with oHW220f and 
            oHW221r from pHW380(8.1) with oHW220f + oHW221r
            Vector: pPD117.01 from the Fire vector (digested with HindIII and 
            EcoRI, ~2.8kb band )
            RE: HindIII and EcoRI digest vector pPD117.01 and Gibson Assembly 
            with insert
FEATURES             Location/Qualifiers
     source          1..5562
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     14..28
                     /locus_tag="oMZ010F_linearize"
                     /label=oMZ010F_linearize
     misc_feature    29..36
                     /locus_tag="FseI"
                     /label=FseI
     misc_feature    37..1987
                     /locus_tag="srj-26p 2kb"
                     /label=srj-26p 2kb
     misc_feature    37..62
                     /locus_tag="oMZ099F"
                     /label=oMZ099F
     misc_feature    complement(1959..1987)
                     /locus_tag="oMZ100R"
                     /label=oMZ100R
     misc_feature    1988..1995
                     /locus_tag="AscI"
                     /label=AscI
     misc_feature    2033..2074
                     /locus_tag="42bp intron"
                     /label=42bp intron
     misc_feature    complement(2063..2081)
                     /locus_tag="oDO025R"
                     /label=oDO025R
     misc_feature    2092..2115
                     /locus_tag="NLS"
                     /label=NLS
     misc_feature    2122..2562
                     /locus_tag="SK GAL4"
                     /label=SK GAL4
     misc_feature    2122..2124
                     /locus_tag="ATG start"
                     /label=ATG start
     misc_feature    2620..2652
                     /locus_tag="VP16 domain"
                     /label=VP16 domain
     misc_feature    2659..2691
                     /locus_tag="VP16 domain(1)"
                     /label=VP16 domain
                     /label=VP16 domain(1)
     misc_feature    2698..2730
                     /locus_tag="VP16 domain(2)"
                     /label=VP16 domain
                     /label=VP16 domain(2)
     misc_feature    2737..2769
                     /locus_tag="VP16 domain(3)"
                     /label=VP16 domain
                     /label=VP16 domain(3)
     misc_feature    2815..3247
                     /locus_tag="let-858 3UTR"
                     /label=let-858 3UTR
                     /label=nonstandard type:  UTR
     misc_feature    complement(3228..3247)
                     /locus_tag="oMZ011R_linearize"
                     /label=oMZ011R_linearize
     CDS             3610..4269
                     /locus_tag="AmpR"
                     /label=AmpR
     rep_origin      4367..5049
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     misc_binding    5389..5411
                     /locus_tag="LacO"
                     /label=LacO
     primer_bind     5396..5415
                     /locus_tag="oDO024F"
                     /label=oDO024F
     primer_bind     5417..5437
                     /locus_tag="M13-rev"
                     /label=M13-rev
     misc_feature    5451..5501
                     /locus_tag="decoy intron"
                     /label=decoy intron
     promoter        join(5502..5562,1..2)
                     /locus_tag="decoy ORF"
                     /label=decoy ORF
ORIGIN
        1 aagcttgcat gcgcggccgc actgactggg ccggcccaag aataaatgct tcgtttgtct
       61 ccctttctgt aaaatacgat gcaaaacgtt aaaaactcgc catttcattc ctgacattcc
      121 tgaaaaacaa gctaattgtt aagagagaaa atgggaaaaa ggaaaaatga gaagagaaag
      181 cgatttttta acgttttgta tcgtattcta cagaaagaga gacaaacaga gcatttttcg
      241 agtaaaaact gtgaaaactt aattatttct catttaaaaa atagttgaac acttttttat
      301 ccaaaaaaac gaacaaaatc gagttaaatc gtggaatttc gaacagaaaa cgaataaaaa
      361 actgcaaatt accgcacgaa acaagcgaga aaaaagcgtg ccgcaggctt gcgttgacgc
      421 tcaagacaaa actttttcaa ataatatatc ctataaggat tatcattatc attaatatta
      481 catcaactgg aatactggta tctttcaaga cacctagcat cattacagcg ggtctgggtg
      541 ctagaatcaa tggaatagtg aaaaccgagc aagtataatc agttgcgaca atccatccgt
      601 gaagaatcaa taggcatcgt ttaactgtga tcatttgcgt gggcgttttg aataaaatac
      661 aaaataatcc gaagacatga attggagctg atacaactga agcgatgtgg aaacttgata
      721 ttaaaaactc aggagtatca atgtatttta tatcaggggt acaagaaaaa tttatttttg
      781 acattggaat aaaatgagta gttgaaaagt ggaggtagtt acttgggcaa aaaatgaaaa
      841 aaaaaattga aaaaccagct gccctcggaa taatgaatat aatattaaaa atggaccaaa
      901 ttcagtgagg aattgtgttt tagaatcgaa catatttttt gcatttttat ttcctatttt
      961 ttaatgtttc catggaaaac attaaaaccc gcatggaatt caaatataaa acaacaacaa
     1021 aaaattgagt gagcaattag tgttgtatat ttacgtgcct gtaaaaaaaa aactgactga
     1081 tcaaatataa ttttaaacat cgaaacaaga atttgttact gaaaattgaa aaaagttttc
     1141 aaaaagtttt tttttctaat ttttttttaa accagctttt cattttgatt ctgataacat
     1201 acaaatcagg gcaaaccgga aaataaattt gatgaatcgc cgaaattttt gacaataatt
     1261 cataagaata tccaaattac gtgtaaaacc ctacagcgag agagagatcg tccaattttt
     1321 gaagttgtcg gaaaaatgtc atgaatatac gcaaatttac cgaaaactca tgctaaattt
     1381 gaaaattcga gttttgaata aaaattagaa agcattatat ataataatag aaaaaaaata
     1441 atttggaatg tttctgaaaa atattgtttt tttttcttga aaatttccca aagatacaca
     1501 tattcaaact gaatttttct tctgtttctg aaaattttga aatcatgcat ttccggcaaa
     1561 aacccgcagt tatgctagtt ttccctccgc cggcaggaga agagagacgc agacagcgag
     1621 agtgtctcgt ttctctgcgc tctctcgaaa tttcaaagtt atcggaaatt tgccatgatt
     1681 atacttattt tgactgggaa aatatgctga acacaaattt ataagactaa tagcttagaa
     1741 aaatgaagtt tttaaaattt caaaaaaaaa aattccattt tatttttcaa aaacccgatt
     1801 actttctata ttttgacgtt tctagaacac agtttataat gtaacaccaa atcatttctc
     1861 agattaatca aattcctcag aattcggaac attcggtcag aagtatacgt cgtagccaaa
     1921 ctcaattacc caaataatct gcacctacat aaaatgctca ttttaccgga aaaagagcac
     1981 ataagctggc gcgcctctag aggatccccg ggattggcca aaggacccaa aggtatgttt
     2041 cgaatgatac taacataaca tagaacattt tcaggaggac ccttggctag gatgggccct
     2101 aaaaagaagc gtaaagctag catgaagctg ttgtcttcaa tggaacaagc ttgcgacata
     2161 tgccggttaa aaaaactcag gtgctcgaag gaaaagccca agtgtgccaa gtgtctgaag
     2221 aacaactcgg agtgccgtta ttctcccaag gctatgaggt ccccactgac cagggcgcat
     2281 ctgacagagg tagaatcaag gttggaaaaa ttggaagatt tgtttctgct catgtttcct
     2341 cgggaaaacc tggacagcat tttgaacatg gactctttgg acgaagtgaa agtaatgttg
     2401 aaacaattat atatgcagga ctatggtgac acggatgctg ctatggatag cttgacccct
     2461 atggaaaatg gcatgtctga agggccgaga aaacacatga agactgcgac atcatcgcca
     2521 aaagtggatg aagatggaag ccaaagtcag ttgagtgtat cgccggaatt cccgctgcag
     2581 tctggtggcg gaggggctcc taggggttcc ggacgggctg acgcattgga cgattttgat
     2641 ctggatatgc tgggaagtga cgccctcgat gattttgacc ttgacatgct tggttcggat
     2701 gcccttgatg actttgacct cgacatgctc ggcagtgacg cccttgatga tttcgacctg
     2761 gacatgctga ttaactaagg taccgaattc gctagccggc catacaagta atccggatga
     2821 tcgacgccaa cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta
     2881 ttatttattt attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa
     2941 atctaaacat ttttttttct ctttccgtct cccaattcgt attccgctcc tctcatctga
     3001 acacaatgtg caagtttatt tatcttctcg ctttcatttc attaggacgt ggggggaatt
     3061 ggtggaaggg ggaaacacac aaaaggatga tggaaatgaa ataaggacac acaatatgca
     3121 acaacattca attcagaaat atggaggaag gtttaaaaga aaacataaaa atatatagag
     3181 gaggaaggaa aactagtaaa aaataagcaa agaaattagg cgaacgatga gaattgtcct
     3241 cgcttgggcc caaaaggcct tttttttttg gcgccgacgt caggtggcac ttttcgggga
     3301 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3361 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3421 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     3481 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     3541 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     3601 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     3661 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     3721 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3781 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3841 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3901 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3961 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4021 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4081 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4141 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4201 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4261 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4321 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4381 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4441 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4501 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4561 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     4621 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     4681 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     4741 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     4801 acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa
     4861 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     4921 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     4981 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5041 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5101 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5161 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     5221 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     5281 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     5341 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     5401 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa
     5461 acatgatctt actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa
     5521 tcactcacaa cgatggatac gctaacaact tggaaatgaa at
//
