LOCUS       Exported                8514 bp DNA     circular SYN 08-AUG-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8514)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 19, 2022 from SnapGene 5.3.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8514
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     204..221
                     /label=M13R (-26)
                     /note="M13R (-26)"
     misc_feature    244..413
                     /label=tol1-L
                     /note="tol1-L"
     enhancer        419..784
                     /note="14xUAS"
     promoter        785..825
                     /note="E1b basal Promoter"
     misc_feature    826..849
                     /label=carp b-actin+24
                     /note="carp b-actin+24"
     intron          join(880..1452,5071..5082)
                     /note="bGlobin intron"
     primer_bind     1497..1524
                     /label=HS 121
     primer_bind     1497..1524
                     /label=RJ0305
                     /note="RJ0305"
     CDS             1501..4338
                     /codon_start=1
                     /label=CaSR
                     /note="CaSR"
                     /translation="MRFHLKFYLHYLVLLGSSCVISTYGPNQRAQKTGDILLGGLFPMH
                     FGVASKDQDLAARPESTECVRYNFRGFRWLQSMIFAIEEINNSSTLLPNITLGYRIFDT
                     CNTVSKALEASLSFVAQNKIDSLNLDEFCNCTGNIPSTIAVVGASGSAVSTAVADLLGL
                     FYIPQISYASSSRLLSNKNQYKSFMRTIPTDEYQAIAMAAIIEHFQWNWVIAIASDDEY
                     GRPGIEKFENEMFHRDICIDLNVLISQYVDEAEIRRLADRIQNSSAKVIVVFASGPDIE
                     PLVKEMVRRNITDRVWLASEAWASSSLVAKPEYLDVMGGTIGFALRAGHIPGFKDFLQQ
                     VHPKKSSHNEFVREFWEETFNCYLEDSPRNADSENGSTSFRPLCTGEEDIASVETPYLD
                     YTHLRISYNVYVAVYAIAQALQDILTCTPGKGLFSNGSCADIRKVEAWQVLKQLRHLNF
                     IDSMGERVRFDNGSELSANYTIINWHRSPEDGSVVFKEVGYYSIHNKNVAKLSIDRSKI
                     LWNGRLTEVPFSNCSVECEPGTRKGIIDGEPTCCFECTECSDGEYSDHKDASFCVKCPN
                     NSWSNGNHTSCFLKEIEFLSWSEPFGIALALFAVLGVLLTAFVLGVFVQFRDTPIVKAS
                     NRELSFLLLFSLICCFSSSLIFIGEPQDWTCRVRQPAFGISFVLCISCILVKTNRVLLV
                     FEAKIPTSLHRKWWGLNLQFLLVFLFTFVQVMICVVWLYNAPPGSYKNYDIDEIIFITC
                     NEGSMMALGFLIGYTCLLAAICFFFAFKSRKLPENFTEAKFITFSMLIFFIVWISFIPA
                     YFSTYGKFVSAVEVIAILASSFSLLACIFFNKVYIILLKPSRNTIEEVRCSTAAHAFKA
                     AAKATLRHSSGFRKRSSSVGGSSASSPSSSISMKTNGNEMESPSTRRHGSKPRVSFGSG
                     TVSLSLSFEEARNSMK"
     variation       3041
                     /label=A>G
                     /note="A>G"
     variation       3271
                     /label=C>G
                     /note="C>G"
     variation       3292
                     /label=A>T
                     /note="A>T"
     primer_bind     3931..3950
                     /label=131
     primer_bind     4339..4374
                     /label=RJ0313
                     /note="RJ0313"
     CDS             4339..4353
                     /codon_start=1
                     /label=GGGGA Linker
                     /note="GGGGA Linker"
                     /translation="GGGGA"
     gene            join(4354..4968,4972..5070)
                     /gene="egfp"
                     /label=egfp
     CDS             4354..5070
                     /codon_start=1
                     /gene="egfp"
                     /product="enhanced green fluorescent protein"
                     /label=EGFP
                     /note="/Contains A206K mutation to ensure monomerization"
                     /db_xref="GI:1377909"
                     /protein_id="AAB02572.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(4467..4485)
                     /label=132
     misc_feature    5083..5543
                     /label=Woodchuck PRE
                     /note="Woodchuck PRE"
     polyA_signal    5551..5971
                     /label=Pout PolyA
                     /note="Pout PolyA"
     misc_feature    5997..6113
                     /label=Tol1-R
                     /note="Tol1-R"
     primer_bind     complement(6118..6135)
                     /label=M13F (-21)
                     /note="M13F (-21)"
     CDS             6713..7573
                     /codon_start=1
                     /label=AmpR
                     /note="AmpR"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atggtccctt tagccagtag cggttctagg cacgggccgt ccgggcggtg gcctggggcg
      301 gaaaactgaa ggggggcggc accggcggct cagcccttgt aatatattat atgcaccact
      361 attggtttac ttatgtcaca gtttgtaagt ttgtaacagc ctgaacctgg atcagcttag
      421 gcctccaagg cggagtactg tcctccgggc tggcggagta ctgtcctccg gcaaggtcgg
      481 agtactgtcc tccgacacta gaggtcggag tactgtcctc cgacgcaagg cggagtactg
      541 tcctccgggc tgcggagtac tgtcctccgg caaggtcgga gtactgtcct ccgacactag
      601 aggtcggagt actgtcctcc gacgcaaggt cggagtactg tcctccgaca ctagaggtcg
      661 gagtactgtc ctccgacgca aggtcggagt actgtcctcc gacactagag gtcggagtac
      721 tgtcctccga cgcaaggcgg agtactgtcc tccgggctgg cggagtactg tcctccggca
      781 agggtcgact ctagagggta tataatggat cccatcgcgt ctcagcctca ctttgagctc
      841 ctccacacga attcccggat cgatcctgag aacttcaggg tgagtttggg gacccttgat
      901 tgttctttct ttttcgctat tgtaaaattc atgttatatg gagggggcaa agttttcagg
      961 gtgttgttta gaatgggaag atgtcccttg tatcaccatg gaccctcatg ataattttgt
     1021 ttctttcact ttctactctg ttgacaacca ttgtctcctc ttattttctt ttcattttct
     1081 gtaacttttt cgttaaactt tagcttgcat ttgtaacgaa tttttaaatt cacttttgtt
     1141 tatttgtcag attgtaagta ctttctctaa tcactttttt ttcaaggcaa tcagggtata
     1201 ttatattgta cttcagcaca gttttagaga acaattgtta taattaaatg ataaggtaga
     1261 atatttctgc atataaattc tggctggcgt ggaaatattc ttattggtag aaacaactac
     1321 atcctggtca tcatcctgcc tttctcttta tggttacaat gatatacact gtttgagatg
     1381 aggataaaat actctgagtc caaaccgggc ccctctgcta accatgttca tgccttcttc
     1441 tttttcctac agctcctggg caacgtggaa ttcgatccat cgattcgaat tcaaggcacc
     1501 atgaggtttc atctgaagtt ttacctgcat tacctggttc tgctgggttc cagctgtgta
     1561 atttcgacct atggcccaaa ccagagggcc cagaagactg gcgatatcct gcttggagga
     1621 cttttcccaa tgcattttgg ggtggcctcc aaagaccagg atcttgcagc acggccagag
     1681 tcaacagagt gtgtcagata taacttccgt ggattccgct ggctgcagtc tatgatcttc
     1741 gctatagagg agatcaataa cagctccact cttcttccaa acattacgct tggctaccgt
     1801 atatttgata cgtgcaacac tgtctcgaaa gcgttggagg cgtcactgag ttttgtcgcg
     1861 cagaataaga ttgattcact gaatctggac gagttctgta actgcacagg gaacatccca
     1921 tcaaccattg cagtggtcgg agcttctgga tctgcagtgt ctacagccgt tgcagacctg
     1981 ctgggtcttt tctacatccc tcagattagc tatgcttcat ccagccgcct tttgagcaac
     2041 aaaaaccagt acaaatcctt catgaggaca atccctacag acgagtacca ggcgattgcc
     2101 atggctgcca tcatcgagca cttccagtgg aactgggtga ttgctatcgc ctctgacgac
     2161 gaatacggcc ggccaggcat cgaaaagttt gagaatgaaa tgtttcaccg ggacatctgc
     2221 attgacctca acgttttaat ttcacagtac gttgatgaag ctgagattcg tcgcctggca
     2281 gatcgcatcc agaactcatc cgccaaggta attgttgtgt ttgctagcgg accagatatc
     2341 gagccgttag ttaaagagat ggtgagacga aacatcacag accgcgtctg gttggcgagc
     2401 gaagcatggg ccagttccag tctagtcgcc aaaccagagt atcttgacgt catgggagga
     2461 accattggct ttgctctgag ggctggacat atacccggct ttaaagactt cttacagcag
     2521 gtccatccaa agaagtcaag ccataacgaa tttgtgcgag aattttggga ggagacgttc
     2581 aactgttatc ttgaagacag cccaaggaat gctgatagtg agaatggcag tacgagtttc
     2641 aggccgctgt gtactgggga agaggacatt gccagtgttg agacgccata tttggactac
     2701 acgcacctca ggatttccta taacgtgtat gtggctgttt atgccatagc acaggcactt
     2761 caggacatac ttacctgcac tccagggaaa ggattgtttt ctaatggctc ttgtgcagat
     2821 attaggaaag ttgaagcatg gcaggttctg aaacaactaa gacacctcaa cttcatagac
     2881 agcatggggg agagagtgcg ctttgacaac ggcagtgagc tttcagccaa ttataccatc
     2941 ataaactggc atcgatcacc cgaggatggg tctgtcgtat tcaaggaggt tggctattac
     3001 agtatacata acaagaacgt ggccaagctt tccattgaca ggagcaaaat cctgtggaac
     3061 ggacgtctaa ctgaggtacc attttccaac tgtagtgtgg aatgtgaacc tggaacaagg
     3121 aaaggaatta ttgatggcga acccacatgt tgctttgagt gtacagaatg ctcggatggg
     3181 gagtacagtg atcataaaga tgccagtttt tgcgttaaat gcccaaacaa ctcctggtcc
     3241 aacggcaatc acacttcttg ctttctgaag gaaatcgagt ttctgtcctg gtccgaaccg
     3301 ttcgggattg cgctggcctt atttgcagtc ctcggggttc tcctaacagc ttttgtgttg
     3361 ggtgtttttg tgcaattccg tgatactcca atcgtgaagg catcaaaccg agagctgtcg
     3421 tttcttttgc ttttctcgct tatatgctgt ttctccagct ctctcatatt cataggcgaa
     3481 ccacaggatt ggacgtgccg ggtacgccaa ccagcattcg gcatcagctt tgtgttatgc
     3541 atctcatgca ttctagtcaa aaccaatcgt gtcctgctag tgttcgaagc taaaatccct
     3601 acaagtctcc accgtaagtg gtggggatta aacctgcagt tcttgctagt cttcctgttc
     3661 acgtttgtgc aggtgatgat ctgcgtggtt tggttgtaca acgctccgcc gggaagttac
     3721 aagaactacg acatcgatga aatcatcttc atcacctgca acgagggctc catgatggct
     3781 ctggggtttc tgattggcta cacgtgcttg ttggccgcca tctgtttttt cttcgcgttc
     3841 aagtctcgga agctgccgga gaatttcacg gaggccaaat tcataacgtt cagcatgctg
     3901 atttttttca tcgtctggat ctcctttatt ccggcgtact tcagcactta cggcaaattc
     3961 gtctcggctg ttgaggtcat agcaatcctg gcttccagct tcagtttgct tgcctgtatc
     4021 tttttcaaca aggtgtatat cattcttctg aaaccgtcga ggaatacgat cgaggaggtg
     4081 cgctgtagca ctgcggctca cgcctttaaa gctgcagcaa aagcgacttt acgacacagt
     4141 tcaggcttta gaaagagatc cagcagtgtt ggcggctcct ccgcatcttc gccctcctca
     4201 tccatcagca tgaagaccaa cgggaatgaa atggagtcac cgtctacgcg gagacacggc
     4261 tcaaaaccga gggtgagctt tggaagtggg acggtcagtc tctccttgag cttcgaggag
     4321 gctagaaact cgatgaaagg aggtggaggt gctgtgagca agggcgagga gctgttcacc
     4381 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     4441 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     4501 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     4561 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4621 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4681 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4741 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4801 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     4861 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4921 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     4981 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     5041 actctcggca tggacgagct gtacaagtaa cctctcgagc ctctagttca ggtgtattgc
     5101 cacaagacaa acatgttaag aaactttccc gttatttacg ctctgttcct gttaatcaac
     5161 ctctggatta caaaatttgt gaaagattga ctgatattct taactatgtt gctcctttta
     5221 cgctgtgtgg atatgctgct ttatagcctc tgtatctagc tattgcttcc cgtacggctt
     5281 tcgttttctc ctccttgtat aaatcctggt tgctgtctct tttagaggag ttgtggcccg
     5341 ttgtccgtca acgtggcgtg gtgtgctctg tgtttgctga cgcaaccccc actggctggg
     5401 gcattgccac cacctgtcaa ctcctttctg ggactttcgc tttccccctc ccgatcgcca
     5461 cggcagaact catcgccgcc tgccttgccc gctgctggac aggggctagg ttgctgggca
     5521 ctgataattc cgtggtgttg tcgtctagta attaagtctc agccaccgtt aactgaacat
     5581 gtcaaaacct gtggagactg ttgagatttg atgttctgaa aagataaagc ctataaataa
     5641 aatgttgccc aaatttcctg cctgatgttt ttctttgtct ttgctacatg gctttgctgc
     5701 tcggatcggc tcactctgtg tatgccacgt tcactttgta ctctccttct cacggtaggt
     5761 ttattatttt tagatgtgca gttagtttct gtgaaataac acaccacaca ctgatattgt
     5821 ctgtgcattg acttggtgag tgcacattgt ttttgatctt gacatattta tatttgattg
     5881 atcaggtgaa ctgtgtgaat ctaaagtgct ccatacagat gttctgcatt gaaaatattc
     5941 tcattttatt agtggaagtg agtgtatgcc agcggccgcg gtaccctgca gtctagacat
     6001 ttcagttgac gaagacaaac aaagttctgt tgtgactatg gggggggggg gcgcctgggg
     6061 atggtctcgc ccggggagta attcagggta gaaccgccac tgcctttagc gacattcact
     6121 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     6181 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     6241 ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac
     6301 gcatctgtgc ggtatttcac accgcatatg gtgcactctc agtacaatct gctctgatgc
     6361 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     6421 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     6481 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     6541 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     6601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     6661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     6721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     6781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     6841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     6901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     6961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     7021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     7081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     7141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     7201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     7261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     7321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     7381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     7441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     7501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     7561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     7621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     7681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     7741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     7801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     7861 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     7921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     7981 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     8041 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     8101 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     8161 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     8221 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     8281 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     8341 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     8401 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     8461 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aaga
//
