LOCUS       Exported                7439 bp ds-DNA     circular SYN 06-MAR-2023
DEFINITION  pUC57.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7439)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 6, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     SA_0_Gal4_noPEST_synCoTC_UAS-mKate2 from 1 to 7439
            SA_1_Gal4_noPEST_synCoTC_UAS-mKate2 from 1 to 7440
            SA_2_Gal4_noPEST_synCoTC_UAS-mKate2 from 1 to 7441
            SA_2_Gal4_noPEST_synCoTC digested with NcoI/SnaBI, mkate2 PCR 
            amplified and inserted using gibson/NEBuilder Hifi
            
            Seq verify with 2061 AvrII_SnaBI_SgH-R and 2086	mKate2-mid-R
FEATURES             Location/Qualifiers
     source          1..7439
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(239..307)
                     /locus_tag="LacZ alpha"
                     /label=LacZ alpha
     primer_bind     364..386
                     /locus_tag="m13/pUC Forward"
                     /label=m13/pUC Forward
     primer_bind     378..395
                     /locus_tag="M13-fwd"
                     /label=M13-fwd
     promoter        414..529
                     /locus_tag="4xnrUAS (Halpern)"
                     /label=4xnrUAS (Halpern)
     promoter        495..511
                     /locus_tag="UAS sequence"
                     /label=UAS sequence
     misc_feature    530..594
                     /locus_tag="E1B"
                     /label=E1B
     misc_feature    557..586
                     /locus_tag="carp beta-actin transcriptional start"
                     /label=carp beta-actin transcriptional start
     5'UTR           607..1295
                     /locus_tag="UBC intron (+1a)"
                     /label=UBC intron (+1a)
     misc_feature    1300..1308
                     /locus_tag="Kozak"
                     /label=Kozak
     misc_feature    join(1303,1306..1309)
                     /locus_tag="Vertebrate Kozak"
                     /label=Vertebrate Kozak
     misc_feature    1306..2004
                     /locus_tag="mKate2 (+2nd Val)"
                     /label=mKate2 (+2nd Val)
     polyA_signal    2036..2227
                     /locus_tag="SV40 late polyA"
                     /label=SV40 late polyA
     misc_feature    2243..2290
                     /locus_tag="48bp_FRT3"
                     /label=48bp_FRT3
     misc_feature    2291..2310
                     /locus_tag="sgH (human ampka1 guide)"
                     /label=sgH (human ampka1 guide)
     misc_feature    2321..2340
                     /locus_tag="sgM site"
                     /label=sgM site
     misc_feature    2344..2391
                     /locus_tag="48bp_FRT"
                     /label=48bp_FRT
     intron          2392..2553
                     /locus_tag="B-actin intron 3'"
                     /label=B-actin intron 3'
     exon            2554..2556
                     /locus_tag="B-actin exon 2 start (SA_0)"
                     /label=B-actin exon 2 start (SA_0)
     misc_feature    2557..2598
                     /locus_tag="V5-tag zf-optim (GKPIPNPLLGLDST)"
                     /label=V5-tag zf-optim (GKPIPNPLLGLDST)
     protein_bind    2605..2658
                     /locus_tag="zf_opt-T2A"
                     /label=zf_opt-T2A
     misc_feature    2659..3312
                     /locus_tag="zf.Gal4-VP16"
                     /label=zf.Gal4-VP16
     protein_bind    2659..3096
                     /locus_tag="zf.Gal4(DBD)"
                     /label=zf.Gal4(DBD)
     protein_bind    3136..3309
                     /locus_tag="zf.VP16 activation domain"
                     /label=zf.VP16 activation domain
     misc_feature    3319..5193
                     /locus_tag="synCoTC"
                     /label=synCoTC
     misc_feature    3319..3403
                     /locus_tag="afp 3'UTR (truncated at polyA)"
                     /label=afp 3'UTR (truncated at polyA)
     terminator      3884..5193
                     /locus_tag="CoTC terminator element"
                     /label=CoTC terminator element
     primer_bind     complement(5214..5234)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     primer_bind     complement(5231..5253)
                     /locus_tag="m13/pUC Reverse"
                     /label=m13/pUC Reverse
     misc_binding    complement(5240..5262)
                     /locus_tag="LacO"
                     /label=LacO
     rep_origin      complement(5602..6284)
                     /direction=LEFT
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     misc_feature    complement(6379..7239)
                     /locus_tag="AmpR gene"
                     /label=AmpR gene
     CDS             complement(6382..7041)
                     /locus_tag="AmpR"
                     /label=AmpR
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggtggc
      421 ttctaatccg tgagtcctag cgggtgacag ccctccgtct tcacaggcgg aggagagtct
      481 tccgtagagt tcctcggagt actgtcctcc gacgcgtgca agggtcgact ctagagggta
      541 tataatggat cccatcgcgt ctcagcctca ctttgagctc ctccacacga attcgatact
      601 aaaagcttgt ttgtgagtat gatgttaaat ataagcaatc gacgtttgtt ttgtcgtttg
      661 ttaaatctag caatgtttta tttcaattaa agtgttaacc ggttcattta aaggtgtatt
      721 tctgaatgag ctaacgtatt agcttttaac gctagccgga ctgtttggtt ttgaggatca
      781 ataaaccaac gtttttttct gatttatttt gctttaaaat ataaaggtat aattcgtcgt
      841 cgagaatttt gggttttgtt cgagtttaga aatttggcgg cgtttttttg taccgcgaaa
      901 cagggcggtg gttcggtgag cttctcaatt catctgcttg acaaacggtt ttcggcaact
      961 gtaaatgcgc tatgatgctt ttattctttt cagcgtgtaa ctttgttaaa tataattgac
     1021 tttccgacta tcgacgttcg tttcatctcc gtgttattgt ttaatggcca ttttctgtgg
     1081 taaactgtcg ccattttgtt ttcagtcgtc cgtttgttcc gcccttactc acagctgtgt
     1141 cagcattagt gatgactgtt ctcacctggc ctgtttgctt tcattattga tgaagagctt
     1201 tctagagaat tctgaagctc gtctttgtcc tgaattcatc tgtattcatt taaaaaaaaa
     1261 aattaacctt aatttttttt tcttctcagc gtaaagcttg ccaccatggt gagcgagctg
     1321 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
     1381 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
     1441 gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
     1501 ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
     1561 cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
     1621 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
     1681 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc
     1741 gagaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc cctgaagctc
     1801 gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa gaaacccgct
     1861 aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag aatcaaggag
     1921 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
     1981 cctagcaaac tggggcacag ataactagta ttaagtctca gccaccgtgt agatccagac
     2041 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2101 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2161 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2221 gttttttcga atgcatctag atgaagttcc tattccgaag ttcctattct tcaaatagta
     2281 taggaacttc gaagatcggc cactacattc tggcctagga ggctgctgcg gttccagagg
     2341 tgggaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt caataaaagt
     2401 taatttaagc gttcattaca ataaattcaa gtgaatctgt tttaagctag cctttaacgt
     2461 ttttaaattc cttcaaacaa attggatcct ctttgcagtt cctcatttct cggtgagctg
     2521 gtgctttaat ccctttactt tttcttcctc tagggaggaa agccaatccc aaaccctctg
     2581 ctgggactgg acagcaccga cgtcgaggga agaggatctc tcctgacctg tggagatgtg
     2641 gaggagaacc ctggcccaat gaaacttcta tcttctatcg aacaggcttg cgacatctgc
     2701 cgcctgaaga agctgaagtg cagcaaggag aagccaaagt gcgccaagtg cctgaagaac
     2761 aactgggagt gcagatactc tcctaagacc aagcgctctc ctctgacccg cgctcacctc
     2821 actgaggtgg agagcagact ggagagactg gagcagctgt ttctcctgat cttcccaaga
     2881 gaggacctgg acatgatcct gaagatggac tctctccagg acatcaaggc tctcctgact
     2941 ggcctgtttg tgcaggacaa cgtgaacaag gatgctgtga ccgacagact ggcttctgtg
     3001 gagaccgaca tgccactgac tctgagacag cacagaatct ctgccacctc ttcttctgag
     3061 gagtctagca acaagggaca gagacagctg actgtgtcta gcagaagcac ccctggaatc
     3121 cagatcagca gagctgctcc tccaactgat gtgtctctcg gagatgagct ccacctggat
     3181 ggagaggatg tggctatggc tcatgctgat gctctggatg actttgacct ggacatgctc
     3241 ggagatggag actctccagg acctggattc acccctcatg actctgctcc ttatggcgct
     3301 ctggacatgt aaactagtat taagtctcag ccaccgttaa ctgaacatgt caaaacctgt
     3361 ggagactgtt gagatttgat gttctgaaaa gataaagcct ataaataaaa caatactcaa
     3421 atacaaaagg tatctttttt aaaaagttag tttactgctt ccttaggctc tttctaaatc
     3481 ccacccacct gaaggacatc aattatggat ttatccttca tgccttgtat aaatacagat
     3541 gtgcatatga atttttatgg gttttttaaa aatgtggttc attctatata cataacaatt
     3601 actttcaaca tacataagcc tacctgtatt tgtgcattat attacatggt acagataaat
     3661 agtcatgtgc cccatgaaca ttttggtcaa agatagatta cagatacaat ggtggtccca
     3721 taagattatg atgccatatt tttactgtac ctcttcttgt gcctagatac acaaatactg
     3781 acttgtgtta taattgcctg cagtattcaa taatatggtg tatagatttg tagcctacga
     3841 gctataacct atactatata gccaaggagt gcaggagggt atgccatcta gatttgtgtg
     3901 agtacactga tgtttgaaca ataaaatcac ctaatgatag gtttctcaaa ccatatccca
     3961 gtccttaagc aacacatgac taatggattc aaccatttcc cctgttcgtt cagattgctt
     4021 tttctttttg cccctacaat tttgttgata atactgtttt ttggggcttt tgtttttgtt
     4081 tttttttttt tttttgagac aagagtctca ctctgtcgtc aggctggagt gcaatggcgc
     4141 tatctcagct cactgcaacc tctgcctccc aggttcaagc gattctcctg cctcagcctc
     4201 ctgagtagct gagattacag gcatgcgcca ccgcgcccgg ctaattttgt atttttagta
     4261 gagacgcagt ttcaccatgt tggtcaggct ggtcttgaac acccgacctc aggtgatcca
     4321 cccgcctcag cttcccaaag tgctgggatt aaaggcgtga gccaccacac ctggccaatt
     4381 tactgtttta taccatgctt ttttgtgtgt attctaggtt ccctaaagca aacatgcatt
     4441 ttaaattttg atagaaccag atgacataca aaagtgcctg ttttctctgc attattaaaa
     4501 ctgaaggtaa tcagtctttt atttctatga ctccaactgg caaaaaatat ttgccatttt
     4561 aacttttttt ttttgagacg gagtcaaaag gtgccagttt tctctgcatt attaaaactg
     4621 aaggtaatca gtcttttatt tctatgactc caactggcaa aagatatttg ccacgttaac
     4681 tttttttttt ttttttttga gacggaggct cacactgtca cccaggctga agtgcagtgg
     4741 catgatctca gctcacttgc aacctccgcc tcccaggttc aagtgattct cctgcctcag
     4801 cctcccaagg agttgggatt acaggcgcat acccggctta ttttttgtat ttttagtaga
     4861 aacgggattt caccgtgtta gccaggatgg tctcaatctc cagacttcgt gatcgcccac
     4921 ctcggcctcc caaagtgctg ggattacagg cgtgagccac cgcgcccggc cctcagtagg
     4981 cgatttttat atgtggtata aagtagaatt cttacagatg gatacccgtt tatgccagtg
     5041 ccatgactga aaccctgctt gtccaacttg aaatatcttc aatgcccatc tattcttcct
     5101 tgatcttgtt ttttcctgtg ataatacaat agtttttacc agttaatcag ttaagatgag
     5161 gcttaccaaa tatatgggag tactactgaa cacgatatca agcttggcgt aatcatggtc
     5221 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     5281 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     5341 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5401 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5461 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5521 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5581 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5641 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5701 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5761 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5821 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5881 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5941 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6001 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     6061 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6121 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6181 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6241 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     6301 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6361 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6421 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6481 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6541 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6601 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6661 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6721 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6781 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6841 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6901 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6961 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     7021 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     7081 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     7141 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     7201 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     7261 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     7321 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     7381 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
