LOCUS       Exported                5240 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 5240)
  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..5240
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     14..28
                     /locus_tag="oMZ010F"
                     /label=oMZ010F
     misc_feature    29..36
                     /locus_tag="FseI"
                     /label=FseI
     misc_feature    37..1665
                     /locus_tag="klp-6p"
                     /label=klp-6p
     misc_feature    1666..1673
                     /locus_tag="AscI"
                     /label=AscI
     misc_feature    1711..1752
                     /locus_tag="42bp intron"
                     /label=42bp intron
     misc_feature    complement(1741..1759)
                     /locus_tag="oDO025R"
                     /label=oDO025R
     misc_feature    1770..1793
                     /locus_tag="NLS"
                     /label=NLS
     misc_feature    1800..2240
                     /locus_tag="SK GAL4"
                     /label=SK GAL4
     misc_feature    1800..1802
                     /locus_tag="ATG start"
                     /label=ATG start
     misc_feature    2298..2330
                     /locus_tag="VP16 domain"
                     /label=VP16 domain
     misc_feature    2337..2369
                     /locus_tag="VP16 domain(1)"
                     /label=VP16 domain
                     /label=VP16 domain(1)
     misc_feature    2376..2408
                     /locus_tag="VP16 domain(2)"
                     /label=VP16 domain
                     /label=VP16 domain(2)
     misc_feature    2415..2447
                     /locus_tag="VP16 domain(3)"
                     /label=VP16 domain
                     /label=VP16 domain(3)
     misc_feature    2493..2925
                     /locus_tag="let-858 3UTR"
                     /label=let-858 3UTR
                     /label=nonstandard type:  UTR
     misc_feature    complement(2906..2925)
                     /locus_tag="oMZ011R"
                     /label=oMZ011R
     CDS             3288..3947
                     /locus_tag="AmpR"
                     /label=AmpR
     rep_origin      4045..4727
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     misc_binding    5067..5089
                     /locus_tag="LacO"
                     /label=LacO
     primer_bind     5074..5093
                     /locus_tag="oDO024F"
                     /label=oDO024F
     primer_bind     5095..5115
                     /locus_tag="M13-rev"
                     /label=M13-rev
     misc_feature    5129..5179
                     /locus_tag="decoy intron"
                     /label=decoy intron
     promoter        join(5180..5240,1..2)
                     /locus_tag="decoy ORF"
                     /label=decoy ORF
ORIGIN
        1 aagcttgcat gcgcggccgc actgactggg ccggccattg tttttttatt gagccttttt
       61 gatgggcatt gtctagtcat tgaaaatatt aacagaaatt ataaaattca ccaaaaaatt
      121 cattaaagca tttttccata tgctttaaaa ttttcaagca taccaaagtt ttcctgtttt
      181 cttgtttcgt tatgggtttt ttagttttca tttagcttac accccccccc cccccccccc
      241 ccccccaacg tcccagacaa tttcaacaaa tcttggttgt gattccgttt caaaacctca
      301 cctatacatt ttctataaat aaagcaaatt atttttaaaa atacctttta tatatatttt
      361 tccttttcca ccctccaatt tatcacattg aaaataaaaa tcataaaacc cccgtttttc
      421 agagtcgacg acttcaataa tttttatttg cttttttaaa attatttttt cgttttgttt
      481 ttttggtcga ataaatctat taaaatggga aatataattg tttttgttca cataacgaga
      541 atttcagaga tgtaggttct ggtcaattcc gtaaatcact gacactccaa agttatgaaa
      601 tgccgcaaag actactgtag gcagtatcaa ttgaatcaat tgttagaaac ctgtttcgag
      661 aagttttgga atattcgtat ttcaacattt tattggctct aaattgttca aaaatctcaa
      721 attttggaaa gtttggtaag ttgccaaaat gactagatac ttttaaatat tgttttttta
      781 ttgagccttt ttgatggatc taacacgttt tggagtttgc tacgatattc gggttttttg
      841 gataattata ttcataagta aaaaatcgga taaatgctgt caaagttgaa ataaactatc
      901 cgaggcttta tcaaaaaatc aattgttttc agaaattctg tgcatttatt caactttttg
      961 ttctctgatt ccaatcttca atcttcatta aatacaactt catacaaatt tttcttcttc
     1021 acatcaaaca ccgtttggtc cgtttccttt cgtcgcttgg agacctacat ggcaacattg
     1081 actccacgtg tatgtagaag aagggtccca ttctccacgc tttactacac tctatactga
     1141 attttccatt tattttcttg tccttttcat tttaaaaaca tttttccgtt cggatttgtt
     1201 tacaaatttt gttctctctg atagaagtat tcaattatta tacttttaat ctgtaaatta
     1261 aaaaaaaaaa ctaaatcctt ttctcatatc acgggaacaa taaatcatga gaatgcgtac
     1321 ttggtgcatt ttatttgacg cgcgatatct cttatcgaaa actacagtaa gagcttaaat
     1381 atctattgtt atttgagttc ttactgtagt tttcgctaag agatatcgcg cgtcaaataa
     1441 aatgcaccaa gtacacattc tcatgattta ttgttcccgt aataccaacc aattattgcg
     1501 acgcaatacc ttcatataca ttttttaatt ttaaaaattt tttttaaaat atagttgaga
     1561 tccttgttgt agtacggtag tagtacgaag atattccaat gtttctttaa aaaaattttc
     1621 caaccaaatt tagttctaaa tttattagtt gaacttttca gaataggcgc gcctctagag
     1681 gatccccggg attggccaaa ggacccaaag gtatgtttcg aatgatacta acataacata
     1741 gaacattttc aggaggaccc ttggctagga tgggccctaa aaagaagcgt aaagctagca
     1801 tgaagctgtt gtcttcaatg gaacaagctt gcgacatatg ccggttaaaa aaactcaggt
     1861 gctcgaagga aaagcccaag tgtgccaagt gtctgaagaa caactcggag tgccgttatt
     1921 ctcccaaggc tatgaggtcc ccactgacca gggcgcatct gacagaggta gaatcaaggt
     1981 tggaaaaatt ggaagatttg tttctgctca tgtttcctcg ggaaaacctg gacagcattt
     2041 tgaacatgga ctctttggac gaagtgaaag taatgttgaa acaattatat atgcaggact
     2101 atggtgacac ggatgctgct atggatagct tgacccctat ggaaaatggc atgtctgaag
     2161 ggccgagaaa acacatgaag actgcgacat catcgccaaa agtggatgaa gatggaagcc
     2221 aaagtcagtt gagtgtatcg ccggaattcc cgctgcagtc tggtggcgga ggggctccta
     2281 ggggttccgg acgggctgac gcattggacg attttgatct ggatatgctg ggaagtgacg
     2341 ccctcgatga ttttgacctt gacatgcttg gttcggatgc ccttgatgac tttgacctcg
     2401 acatgctcgg cagtgacgcc cttgatgatt tcgacctgga catgctgatt aactaaggta
     2461 ccgaattcgc tagccggcca tacaagtaat ccggatgatc gacgccaacg tcgttgaatt
     2521 ttcaaatttt aaatactgaa tatttgtttt ttttcctatt atttatttat tctctttgtg
     2581 ttttttttct tgctttctaa aaaattaatt caatccaaat ctaaacattt ttttttctct
     2641 ttccgtctcc caattcgtat tccgctcctc tcatctgaac acaatgtgca agtttattta
     2701 tcttctcgct ttcatttcat taggacgtgg ggggaattgg tggaaggggg aaacacacaa
     2761 aaggatgatg gaaatgaaat aaggacacac aatatgcaac aacattcaat tcagaaatat
     2821 ggaggaaggt ttaaaagaaa acataaaaat atatagagga ggaaggaaaa ctagtaaaaa
     2881 ataagcaaag aaattaggcg aacgatgaga attgtcctcg cttgggccca aaaggccttt
     2941 tttttttggc gccgacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3001 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3061 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3121 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3181 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3241 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3301 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     3361 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3421 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3481 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3541 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     3601 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     3661 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     3721 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     3781 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     3841 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3901 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3961 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     4021 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4081 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     4141 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     4201 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     4261 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     4321 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     4381 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     4441 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     4501 tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     4561 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     4621 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     4681 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     4741 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     4801 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     4861 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     4921 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     4981 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     5041 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     5101 acagctatga ccatgattac gccaagctgt aagtttaaac atgatcttac taactaacta
     5161 ttctcattta aattttcaga gcttaaaaat ggctgaaatc actcacaacg atggatacgc
     5221 taacaacttg gaaatgaaat
//
