LOCUS       Exported                7958 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    TRPC3-Myc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7958)
  AUTHORS   Li HS, Xu XZ, Montell C
  TITLE     Activation of a TRPC3-dependent cation current through the 
            neurotrophin BDNF.
  JOURNAL   Neuron. 1999 Sep . 24(1):261-73.
  PUBMED    10677043
REFERENCE   2  (bases 1 to 7958)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7958
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3229..3258
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     promoter        complement(3277..3295)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3278..3295)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3315..3332)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3321..3545
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3591..4019
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3678..3697)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3888..3909
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4028..4048)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4033..4362
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4213..4348
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4275..4294
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4429..5223
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4483..4502)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5093..5112
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5397..5518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5434..5453)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5488..5507
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5567..5583)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5567..5583)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5580..5602)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5591..5607
                     /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(5615..5645)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5660..5681
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5798..5815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5969..6557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6049..6068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6728..7588)
                     /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"
     primer_bind     7351..7370
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7589..7693)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7768..7787)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac catggaggga agcccatccc tgagacgcat gacagtgatg
      721 cgggagaagg gccggcgcca ggctgtcagg ggcccggcct tcatgttcaa tgaccgcggc
      781 accagcctca ccgccgagga ggagcgcttc ctcgacgccg ccgagtacgg caacatccca
      841 gtggtgcgca agatgctgga ggagtccaag acgctgaacg tcaactgcgt ggactacatg
      901 ggccagaacg cgctgcagct ggctgtgggc aacgagcacc tggaggtgac cgagctgctg
      961 ctcaagaagg agaacctggc gcgcattggc gacgccctgc tgctcgccat cagcaagggc
     1021 tacgtgcgca tcgtagaggc catcctcaac caccctggct tcgcggccag caagcgtctc
     1081 actctgagcc cctgtgagca ggagctgcag gacgacgact tctacgctta cgacgaggac
     1141 ggcacgcgct tctcgccgga catcaccccc atcatcctgg cggcgcactg ccagaaatac
     1201 gaagtggtgc acatgctgct gatgaagggt gccaggatcg agcggccgca cgactatttc
     1261 tgcaagtgcg gggactgcat ggagaagcag aggcacgact ccttcagcca ctcacgctcg
     1321 aggatcaatg cctacaaggg gctggccagc ccggcttacc tctcattgtc cagcgaggac
     1381 ccggtgctta cggccctaga gctcagcaac gagctggcca agctggccaa catagagaag
     1441 gagttcaaga atgactatcg gaagctctcc atgcaatgca aagactttgt agtgggtgtg
     1501 ctggatctct gccgagactc agaagaggta gaagccattc tgaatggaga tctggaatca
     1561 gcagagcctc tggaggtaca caggcacaaa gcttcattaa gtcgtgtcaa acttgccatt
     1621 aagtatgaag tcaaaaagtt tgtggctcat cccaactgcc agcagcagct cttgacgatc
     1681 tggtatgaga acctctcagg cctaagggag cagaccatag ctatcaagtg tctcgttgtg
     1741 ctggtcgtgg ccctgggcct tccattcctg gccattggct actggatcgc accttgcagc
     1801 aggctgggga aaattctgcg aagccctttt atgaagtttg tagcacatgc agcttctttc
     1861 atcatcttcc tgggtctgct tgtgttcaat gcctcagaca ggttcgaagg catcaccacg
     1921 ctgcccaata tcacagttac tgactatccc aaacagatct tcagggtgaa aaccacccag
     1981 tttacatgga ctgaaatgct aattatggtc tgggttcttg gaatgatgtg gtctgaatgt
     2041 aaagagctct ggctggaagg acctagggaa tacattttgc agttgtggaa tgtgcttgac
     2101 tttgggatgc tgtccatctt cattgctgct ttcacagcca gattcctagc tttccttcag
     2161 gcaacgaagg cacaacagta tgtggacagt tacgtccaag agagtgacct cagtgaagtg
     2221 acactcccac cagagataca gtatttcact tatgctagag ataaatggct cccttctgac
     2281 cctcagatta tatctgaagg cctttatgcc atagctgttg tgctcagctt ctctcggatt
     2341 gcgtacatcc tccctgcaaa tgagagcttt ggccccctgc agatctctct tggaaggact
     2401 gtaaaggaca tattcaagtt catggtcctc tttattatgg tgttttttgc ctttatgatt
     2461 ggcatgttca tactttattc ttactacctt ggggctaaag ttaatgctgc ttttaccact
     2521 gtagaagaaa gtttcaagac tttattttgg tcaatatttg ggttgtctga agtgacttcc
     2581 gttgtgctca aatatgatca caaattcata gaaaatattg gatacgttct ttatggaata
     2641 tacaatgtaa ctatggtggt cgttttactc aacatgctaa ttgctatgat taatagctca
     2701 tatcaagaaa ttgaggatga cagtgatgta gaatggaagt ttgctcgttc aaaactttgg
     2761 ttatcctatt ttgatgatgg aaaaacatta cctccacctt tcagtctagt tcctagtcca
     2821 aaatcatttg tttatttcat catgcgaatt gttaactttc ccaaatgcag aaggagaagg
     2881 cttcagaagg atatagaaat gggaatgggt aactcaaagt ccaggttaaa cctcttcact
     2941 cagtctaact caagagtttt tgaatcacac agttttaaca gcattctcaa tcagccaaca
     3001 cgttatcagc agataatgaa aagacttata aagcggtatg ttttgaaagc acaagtagac
     3061 aaagaaaatg atgaagttaa tgaaggtgaa ttaaaagaaa tcaagcaaga tatctccagc
     3121 cttcgttatg aacttttgga agacaagagc caagcaactg aggaattagc cattctaatt
     3181 cataaactta gtgagaaact gaatcccagc atgctgagat gtgaactcga gcagaaactc
     3241 atctctgaag aggatctgta atctagaggg ccctattcta tagtgtcacc taaatgctag
     3301 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3361 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3421 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3481 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3541 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3601 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3661 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3721 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3781 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3841 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3901 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3961 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4021 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4081 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4141 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4201 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4261 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4321 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     4381 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     4441 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     4501 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     4561 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     4621 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     4681 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     4741 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     4801 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     4861 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     4921 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     4981 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5041 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5101 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5161 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5221 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     5281 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     5341 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     5401 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5461 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5521 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     5581 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     5641 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     5701 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     5761 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5821 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5881 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5941 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6001 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6061 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6121 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6181 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6241 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6301 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6361 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6421 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6481 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6541 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6601 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6661 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6721 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6781 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6841 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6901 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6961 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7021 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7081 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7141 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7201 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7261 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7321 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7381 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7441 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7501 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     7561 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     7621 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7681 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     7741 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     7801 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     7861 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     7921 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
