LOCUS       Exported                6007 bp ds-DNA     circular SYN 29-AUG-2019
DEFINITION  Vector that allows high-level transient expression in vertebrate 
            cells as well as in vitro transcription/translation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6007)
  AUTHORS   I.M.A.G.E. Consortium
  TITLE     Direct Submission
  JOURNAL   Exported Aug 29, 2019 from SnapGene Viewer 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6007
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          88..2031
                     /organism="Danio rerio"
                     /strain="AB"
                     /mol_type="mRNA"
                     /dev_stage="tailbud"
                     /db_xref="taxon:7955"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             88..2031
                     /codon_start=1
                     /gene="npas4l"
                     /allele="wildtype"
                     /product="Npas4l"
                     /label=npas4l
                     /note="bHLH-PAS transcription factor"
                     /protein_id="ANP24257.1"
                     /translation="MSCGDSGIRRILRASKRFRSTKGASKARRDQMNSEIRNLRALLPI
                     SPEHRLSYLHSMSITCTYIRKSVELRGVCEESTVFSAVNGCVPQDCALQDCALQECVLQ
                     ECVLQECVLQECVLQALPGFIVAFTTDGKLLYVSENVHEYLGLSMVDVLQSDSFFDMLD
                     RSDVEAVRSVLADASPSGERWVVCRMLVSKAMRLRSSCCPLLVRIRLRDGVCVSLCRPT
                     ADRLPARNADFHTHHSADMRLASASSSVLFHLGFSADELIGRSWYELLHPDDLRHAADR
                     HAAILAAATADAEMLIRVQCKDLSWVWMYTHASATAERDAISCSNYMISEAEAVYLQQR
                     LSSSSSSSTSSSSSSASPQCSSVSDSSDAHSAHTLFCTPPYSPTSSQCSDFLSEGYGSL
                     EALVDSAFCSPPYPPLFPNPCCTPTVCPPDTALDPARLCPPDSALVPAALCPPDAALDP
                     TQLCPPGAALDPTLLCPPGAALDPTQLCLPDGALVPAVFCPPNGALVPARLCPSDAALV
                     PAALGRPDAALVPVCRPLQVCECPLDGAVPLEDLSMFPLPQGGGSRLMPPEASPTTHTH
                     FSYDAAQQAGIGTLAMQIHTLICSFDAYSAARHTHSHHTSCWAPEPLLDEGIIDSILRE
                     LDSTHTDAHTLI"
     gene            88..2031
                     /gene="npas4l"
                     /allele="wildtype"
                     /label=npas4l
                     /note="cloche"
     misc_feature    88..143
                     /label=Exon1
     primer_bind     complement(137..156)
                     /label=npas4l_bHLHless_rev
     misc_feature    144..303
                     /label=Exon2
     primer_bind     complement(147..168)
                     /label=npas4l_binding_mut_rev
     misc_feature    157..213
                     /label=Helix (bHLH)
     misc_feature    169..177
                     /label=DNA-binding motif?
     primer_bind     178..202
                     /label=npas4l_binding_mut_fwd
     misc_feature    214..243
                     /label=loop
     misc_feature    244..288
                     /label=Helix (bHLH#)
     primer_bind     289..316
                     /label=npas4l_bHLHless_fwd2
     misc_feature    304..536
                     /label=Exon3
     primer_bind     352..369
                     /label=npas4l qPCR_fwd (published)
     primer_bind     364..384
                     /label=npas4l qPCR_fwd (published)
     primer_bind     458..478
                     /label=npas4l-qPCR3_fwd
     primer_bind     complement(490..511)
                     /label=npas4l qPCR_rev (published)
     misc_feature    537..631
                     /label=Exon4
     misc_feature    632..833
                     /label=Exon5
     primer_bind     complement(645..664)
                     /label=npas4l-qPCR3_rev
     misc_feature    834..943
                     /label=Exon6
     misc_feature    944..1076
                     /label=Exon7
     primer_bind     1061..1084
                     /label=npas4l-qPCR2_fwd
     misc_feature    1077..2031
                     /label=Exon8
     misc_feature    1132
                     /label=t in the genomic assembly (SNP?)
     primer_bind     complement(1291..1310)
                     /label=npas4l-qPCR2_rev
     misc_feature    2032..2056
                     /label=stop codons
                     /note="stop codons in all three reading frames"
     promoter        complement(2034..2051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2056..2190
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2303..2321)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2342..2358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2366..2382
                     /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(2390..2420)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2435..2456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2744..3332)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3503..4363)
                     /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"
     promoter        complement(4364..4468)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4494..4949)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5023..6007
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccaccatg agctgcggag actcgggcat caggaggatc
      121 ctccgcgcgt ccaagcgctt caggtccacc aaaggagcat ctaaggcgcg gcgtgatcag
      181 atgaacagtg agatacggaa cctgcgagcg ctgctgccca tcagcccaga gcacagactc
      241 tcatacctgc acagcatgag catcacctgc acctacatca ggaagagtgt ggagctgaga
      301 ggtgtgtgtg aggaatccac agttttttct gccgtgaatg gatgtgtccc gcaggactgt
      361 gctctacagg actgtgctct acaggagtgt gttctgcagg agtgtgttct gcaggagtgt
      421 gttctgcagg agtgtgttct gcaggctctg ccggggttta tcgttgcctt taccacagat
      481 ggaaaactgc tgtatgtgtc tgagaacgta cacgagtatc tgggtctgtc catggtggac
      541 gttctccaga gcgactcttt cttcgatatg ctggaccgca gtgatgtgga ggcggtgagg
      601 agtgtgttag ctgacgccag tccttcagga gagcggtggg ttgtgtgtcg tatgctggtg
      661 tctaaagcga tgcgtctgcg gagctcctgc tgccccctgc tggtgcggat ccgtctgcgg
      721 gacggagtgt gtgtgtcgct ctgcagaccc accgctgacc ggctgccggc gcgcaacgct
      781 gacttccaca cacaccacag cgcagacatg aggctcgcca gcgcatcctc cagtgtgttg
      841 ttccacctgg gcttctcagc ggacgagctg attggtcgat cctggtacga gctcctgcac
      901 cccgacgacc tccgacacgc tgctgaccgc cacgccgcaa tccttgcagc cgccacagca
      961 gacgcagaga tgctgatccg ggttcagtgt aaggatctgt cctgggtgtg gatgtacaca
     1021 cacgcatcag caacagcaga gcgggacgcg atcagctgct ccaactacat gatcagtgaa
     1081 gcagaggcag tgtatctgca gcagagactc tcctcctcct cctcctcctc cacctcctcc
     1141 tcctcctcct ctgcctctcc tcagtgtagc tccgtctcgg acagcagtga tgcccacagc
     1201 gcacacacac tcttctgcac gccgccctac agccccactt cctctcagtg ctctgacttc
     1261 ctgtctgaag gttatggctc tctggaggcg ctggtggatt ctgccttctg ctctccgccg
     1321 tacccgccgc tgttccccaa cccatgctgc acccccactg tctgcccacc ggacactgcc
     1381 ctggaccccg ctcggctctg cccgccagat tctgccctgg tccccgctgc gctctgtccg
     1441 ccggacgctg ccctggaccc cactcagctc tgcccgccgg gagctgccct ggaccccact
     1501 ctgctctgcc cgccgggagc tgccctggac cccactcagc tctgcctgcc ggacggtgcc
     1561 ttggtccctg ctgtgttctg cccgccgaac ggtgccctgg tccctgcccg gctctgcccg
     1621 tcagatgctg ccctggtccc cgctgcactc ggtcggccgg acgctgccct ggtgcctgtg
     1681 tgtcggccac tgcaggtgtg tgagtgccct ctggatggcg ctgtgccgct ggaggacctc
     1741 agcatgttcc cgctgccgca gggcggaggc tctcgtctga tgccgcctga agcatcaccc
     1801 actacccaca cacacttctc gtatgatgca gcacagcagg cggggatcgg cacgttggca
     1861 atgcagatac acacactcat ctgcagcttc gatgcgtaca gcgccgcacg acacacacac
     1921 tctcaccaca cctcctgctg ggcaccagag ccactgctgg acgagggcat catcgacagc
     1981 atcctcagag agctggacag cacacacact gatgcacaca cactgatcta gaactatagt
     2041 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     2101 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2161 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2221 caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg
     2281 gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     2341 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     2401 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2461 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2521 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2581 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2641 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2701 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2761 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2821 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2881 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2941 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3001 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3061 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3121 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3181 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3241 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3301 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     3361 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3421 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3481 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3541 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3601 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3661 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3721 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3781 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3841 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3901 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3961 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4021 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4081 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4141 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4201 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     4261 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     4321 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     4381 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4441 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     4501 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4561 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4621 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4681 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4741 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4801 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4861 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4921 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     4981 ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccatag ccaattcaat
     5041 atggcgtata tggactcatg ccaattcaat atggtggatc tggacctgtg ccaattcaat
     5101 atggcgtata tggactcgtg ccaattcaat atggtggatc tggaccccag ccaattcaat
     5161 atggcggact tggcaccatg ccaattcaat atggcggact tggcactgtg ccaactgggg
     5221 aggggtctac ttggcacggt gccaagtttg aggaggggtc ttggccctgt gccaagtccg
     5281 ccatattgaa ttggcatggt gccaataatg gcggccatat tggctatatg ccaggatcaa
     5341 tatataggca atatccaata tggccctatg ccaatatggc tattggccag gttcaatact
     5401 atgtattggc cctatgccat atagtattcc atatatgggt tttcctattg acgtagatag
     5461 cccctcccaa tgggcggtcc catataccat atatggggct tcctaatacc gcccatagcc
     5521 actcccccat tgacgtcaat ggtctctata tatggtcttt cctattgacg tcatatgggc
     5581 ggtcctattg acgtatatgg cgcctccccc attgacgtca attacggtaa atggcccgcc
     5641 tggctcaatg cccattgacg tcaataggac cacccaccat tgacgtcaat gggatggctc
     5701 attgcccatt catatccgtt ctcacgcccc ctattgacgt caatgacggt aaatggccca
     5761 cttggcagta catcaatatc tattaatagt aacttggcaa gtacattact attggaagga
     5821 cgccagggta cattggcagt actcccattg acgtcaatgg cggtaaatgg cccgcgatgg
     5881 ctgccaagta catccccatt gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc
     5941 attgacgtaa atgggcggta ggcgtgccta atgggaggtc tatataagca atgctcgttt
     6001 agggaac
//
