LOCUS       Exported                7441 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 7441)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Mar 6, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     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..7441
                     /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..2558
                     /locus_tag="B-actin exon 2 start (SA_2)"
                     /label=B-actin exon 2 start (SA_2)
     misc_feature    2559..2600
                     /locus_tag="V5-tag zf-optim (GKPIPNPLLGLDST)"
                     /label=V5-tag zf-optim (GKPIPNPLLGLDST)
     protein_bind    2607..2660
                     /locus_tag="zf_opt-T2A"
                     /label=zf_opt-T2A
     misc_feature    2661..3314
                     /locus_tag="zf.Gal4-VP16"
                     /label=zf.Gal4-VP16
     protein_bind    2661..3098
                     /locus_tag="zf.Gal4(DBD)"
                     /label=zf.Gal4(DBD)
     protein_bind    3138..3311
                     /locus_tag="zf.VP16 activation domain"
                     /label=zf.VP16 activation domain
     misc_feature    3321..5195
                     /locus_tag="synCoTC"
                     /label=synCoTC
     misc_feature    3321..3405
                     /locus_tag="afp 3'UTR (truncated at polyA)"
                     /label=afp 3'UTR (truncated at polyA)
     terminator      3886..5195
                     /locus_tag="CoTC terminator element"
                     /label=CoTC terminator element
     primer_bind     complement(5216..5236)
                     /locus_tag="M13-rev"
                     /label=M13-rev
     primer_bind     complement(5233..5255)
                     /locus_tag="m13/pUC Reverse"
                     /label=m13/pUC Reverse
     misc_binding    complement(5242..5264)
                     /locus_tag="LacO"
                     /label=LacO
     rep_origin      complement(5604..6286)
                     /direction=LEFT
                     /locus_tag="ColE1 origin"
                     /label=ColE1 origin
     misc_feature    complement(6381..7241)
                     /locus_tag="AmpR gene"
                     /label=AmpR gene
     CDS             complement(6384..7043)
                     /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 tagggggagg aaagccaatc ccaaaccctc
     2581 tgctgggact ggacagcacc gacgtcgagg gaagaggatc tctcctgacc tgtggagatg
     2641 tggaggagaa ccctggccca atgaaacttc tatcttctat cgaacaggct tgcgacatct
     2701 gccgcctgaa gaagctgaag tgcagcaagg agaagccaaa gtgcgccaag tgcctgaaga
     2761 acaactggga gtgcagatac tctcctaaga ccaagcgctc tcctctgacc cgcgctcacc
     2821 tcactgaggt ggagagcaga ctggagagac tggagcagct gtttctcctg atcttcccaa
     2881 gagaggacct ggacatgatc ctgaagatgg actctctcca ggacatcaag gctctcctga
     2941 ctggcctgtt tgtgcaggac aacgtgaaca aggatgctgt gaccgacaga ctggcttctg
     3001 tggagaccga catgccactg actctgagac agcacagaat ctctgccacc tcttcttctg
     3061 aggagtctag caacaaggga cagagacagc tgactgtgtc tagcagaagc acccctggaa
     3121 tccagatcag cagagctgct cctccaactg atgtgtctct cggagatgag ctccacctgg
     3181 atggagagga tgtggctatg gctcatgctg atgctctgga tgactttgac ctggacatgc
     3241 tcggagatgg agactctcca ggacctggat tcacccctca tgactctgct ccttatggcg
     3301 ctctggacat gtaaactagt attaagtctc agccaccgtt aactgaacat gtcaaaacct
     3361 gtggagactg ttgagatttg atgttctgaa aagataaagc ctataaataa aacaatactc
     3421 aaatacaaaa ggtatctttt ttaaaaagtt agtttactgc ttccttaggc tctttctaaa
     3481 tcccacccac ctgaaggaca tcaattatgg atttatcctt catgccttgt ataaatacag
     3541 atgtgcatat gaatttttat gggtttttta aaaatgtggt tcattctata tacataacaa
     3601 ttactttcaa catacataag cctacctgta tttgtgcatt atattacatg gtacagataa
     3661 atagtcatgt gccccatgaa cattttggtc aaagatagat tacagataca atggtggtcc
     3721 cataagatta tgatgccata tttttactgt acctcttctt gtgcctagat acacaaatac
     3781 tgacttgtgt tataattgcc tgcagtattc aataatatgg tgtatagatt tgtagcctac
     3841 gagctataac ctatactata tagccaagga gtgcaggagg gtatgccatc tagatttgtg
     3901 tgagtacact gatgtttgaa caataaaatc acctaatgat aggtttctca aaccatatcc
     3961 cagtccttaa gcaacacatg actaatggat tcaaccattt cccctgttcg ttcagattgc
     4021 tttttctttt tgcccctaca attttgttga taatactgtt ttttggggct tttgtttttg
     4081 tttttttttt tttttttgag acaagagtct cactctgtcg tcaggctgga gtgcaatggc
     4141 gctatctcag ctcactgcaa cctctgcctc ccaggttcaa gcgattctcc tgcctcagcc
     4201 tcctgagtag ctgagattac aggcatgcgc caccgcgccc ggctaatttt gtatttttag
     4261 tagagacgca gtttcaccat gttggtcagg ctggtcttga acacccgacc tcaggtgatc
     4321 cacccgcctc agcttcccaa agtgctggga ttaaaggcgt gagccaccac acctggccaa
     4381 tttactgttt tataccatgc ttttttgtgt gtattctagg ttccctaaag caaacatgca
     4441 ttttaaattt tgatagaacc agatgacata caaaagtgcc tgttttctct gcattattaa
     4501 aactgaaggt aatcagtctt ttatttctat gactccaact ggcaaaaaat atttgccatt
     4561 ttaacttttt ttttttgaga cggagtcaaa aggtgccagt tttctctgca ttattaaaac
     4621 tgaaggtaat cagtctttta tttctatgac tccaactggc aaaagatatt tgccacgtta
     4681 actttttttt tttttttttt gagacggagg ctcacactgt cacccaggct gaagtgcagt
     4741 ggcatgatct cagctcactt gcaacctccg cctcccaggt tcaagtgatt ctcctgcctc
     4801 agcctcccaa ggagttggga ttacaggcgc atacccggct tattttttgt atttttagta
     4861 gaaacgggat ttcaccgtgt tagccaggat ggtctcaatc tccagacttc gtgatcgccc
     4921 acctcggcct cccaaagtgc tgggattaca ggcgtgagcc accgcgcccg gccctcagta
     4981 ggcgattttt atatgtggta taaagtagaa ttcttacaga tggatacccg tttatgccag
     5041 tgccatgact gaaaccctgc ttgtccaact tgaaatatct tcaatgccca tctattcttc
     5101 cttgatcttg ttttttcctg tgataataca atagttttta ccagttaatc agttaagatg
     5161 aggcttacca aatatatggg agtactactg aacacgatat caagcttggc gtaatcatgg
     5221 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5281 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5341 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5401 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7381 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7441 c
//
