LOCUS       Exported                7128 bp ds-DNA     circular SYN 14-AUG-2017
DEFINITION  Cloning vector pCLIPf-NK1R, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7128)
  AUTHORS   Sun L, Provost C, Ghosh I, Xu MQ.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-APR-2011) Research Department, New England Biolabs, 
            240 County Road, Ipswich, MA 01938, USA
REFERENCE   2  (bases 1 to 7128)
  AUTHORS   Addgene
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Aug 14, 2017 from SnapGene 3.1.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7128
                     /organism="Cloning vector pCLIPf-NK1R"
                     /mol_type="other DNA"
     promoter        251..818
                     /note="CMV immediate early promoter region"
     promoter        863..880
                     /note="T7 promoter (clockwise)"
     gene            921..2816
                     /gene="CLIP10f/NK1R"
                     /product="O-6-methylguanine-DNA methyltransferase / NK1 
                     receptor fusion with N-terminal ER signal and C-terminal 
                     FLAG epitope and His-tag"
                     /note="CLIP10f/NK1R"
                     /note="5-hydroxytryptamine receptor 3A (5HT3R) signal 
                     sequence 921..992, CLIP10f sequence 999..1544, truncated 
                     NK1 receptor sequence 1551..2729, FLAG epitope sequence 
                     2730..2753, polyhistidine tag sequence 2784..2813; CLIP10f 
                     optimized for mammalian expression; possible alternative 
                     start 774"
     misc_RNA        3212..3794
                     /note="internal ribosome entry site (IRES) from 
                     encephalomyocarditis virus (ECMV); allows polycistronic 
                     expression of CLIP10f/NK1R and neoR, and required for 
                     expression of neoR"
     CDS             3815..4618
                     /codon_start=1
                     /gene="aph(3')-II"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="aph(3')-II"
                     /note="neoR, kanR, nptII (confers resistance to neomycin 
                     and kanamycin)"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3815..4618
                     /gene="aph(3')-II"
                     /note="aph(3')-II"
     promoter        complement(5020..5049)
                     /note="Plac promoter (-35 signal TTTACA, -10 signal TATGTT)
                     (counter-clockwise)"
     rep_origin      complement(5373..5961)
                     /direction=LEFT
                     /note="pUC19 origin of replication (counter-clockwise) 
                     (RNAII -35 to RNA/DNA switch point)"
     CDS             complement(6132..6992)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="bla"
                     /note="ampR (confers resistance to ampicillin)"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6132..6992)
                     /gene="bla"
                     /note="bla"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatcacc atgcggctct gcatcccgca ggtgctgttg gccttgttcc
      961 tttccatgct gacagggccg ggagaaggca gcgctagcat ggacaaagac tgcgaaatga
     1021 agcgcaccac cctggatagc cctctgggca agctggaact gtctgggtgc gaacagggcc
     1081 tgcaccgtat catcttcctg ggcaaaggaa catctgccgc cgacgccgtg gaagtgcctg
     1141 ccccagccgc cgtgctgggc ggaccagagc cactgatcca ggccaccgcc tggctcaacg
     1201 cctactttca ccagcctgag gccatcgagg agttccctgt gccagccctg caccacccag
     1261 tgttccagca ggagagcttt acccgccagg tgctgtggaa actgctgaaa gtggtgaagt
     1321 tcggagaggt catcagcgag agccacctgg ccgccctggt gggcaatccc gccgccaccg
     1381 ccgccgtgaa caccgccctg gacggaaatc ccgtgcccat tctgatcccc tgccaccggg
     1441 tggtgcaggg cgacagcgac gtggggccct acctgggcgg gctcgccgtg aaagagtggc
     1501 tgctggccca cgagggccac agactgggca agcctgggct gggtcctgca ggttccacta
     1561 acacctcgga acccaatcag ttcgtgcaac cagcctggca aattgtcctt tgggcagctg
     1621 cctacacggt cattgtggtg acctctgtgg tgggcaacgt ggtagtgatg tggatcatct
     1681 tagcccacaa aagaatgagg acagtgacga actattttct ggtgaacctg gccttcgcgg
     1741 aggcctccat ggctgcattc aatacagtgg tgaacttcac ctatgctgtc cacaacgaat
     1801 ggtactacgg cctgttctac tgcaagttcc acaacttctt tcccatcgcc gctgtcttcg
     1861 ccagtatcta ctccatgacg gctgtggcct ttgataggta catggccatc atacatcccc
     1921 tccagccccg gctgtcagcc acagccacca aagtggtcat ctgtgtcatc tgggtcctgg
     1981 ctctcctgct ggccttcccc cagggctact actcaaccac agagaccatg cccagcagag
     2041 tcgtgtgcat gatcgaatgg ccagagcatc cgaacaagat ttatgagaaa gtgtaccaca
     2101 tctgtgtgac tgtgctgatc tacttcctcc ccctgctggt gattggctat gcatacaccg
     2161 tagtgggaat cacactatgg gccagtgaga tccccgggga ctcctctgac cgctaccacg
     2221 agcaagtctc tgccaagcgc aaggtggtca aaatgatgat tgtcgtggtg tgcaccttcg
     2281 ccatctgcag gctgcccttc cacatcttct tcctcctgcc ctacaccaac ccagatctct
     2341 acctgaagaa gtttatccag caggtctacc tggccatcat gtggctggcc atgagctcca
     2401 ccatgtacaa ccccatcatc tactgctgcc tcaatgacag gttccgtctg ggcttcaagc
     2461 atgccttccg gtgctgcccc ttcatcagcg ccggcgacta tgaggggctg gaaatgaaat
     2521 ccacccggta tctccagacc cagggcagtg tgtacaaagt cagccgcctg gagaccacca
     2581 tctccacagt ggtgggggcc cacgaggagg agccagagga cggccccaag gccacaccct
     2641 cgtccctgga cctgacctcc aactgctctt cacgaagtga ctccaagacc atgacagaga
     2701 gcttcagctt ctcctccaat gtgctctccg actacaagga cgacgatgac aagaccggtg
     2761 tgccgcgcgg cagcggcagc agccatcatc atcatcatca tcaccatcac cattaaggat
     2821 cctgactcga gtgaggcggc cgcatagata actgatccag tgtgctggaa ttaattcgct
     2881 gtctgcgagg gccagctgtt ggggtgagta ctccctctca aaagcgggca tgacttctgc
     2941 gctaagattg tcagtttcca aaaacgagga ggatttgata ttcacctggc ccgcggtgat
     3001 gcctttgagg gtggccgcgt ccatctggtc agaaaagaca atctttttgt tgtcaagctt
     3061 gaggtgtggc aggcttgaga tctggccata cacttgagtg acaatgacat ccactttgcc
     3121 tttctctcca caggtgtcca ctcccaggtc caactgcagg tcgagcatgc atctagggcg
     3181 gccaattccg cccctctccc cccccccctt ttccctcccc cccccctaac gttactggcc
     3241 gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc accatattgc
     3301 cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg agcattccta
     3361 ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag
     3421 ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc aggcagcgga
     3481 accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa gatacacctg
     3541 caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa agagtcaaat
     3601 ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta ccccattgta
     3661 tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg aggttaaaaa
     3721 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataag
     3781 cttgccacaa cccgggataa ttcctgcagc caatatggga tcggccattg aacaagatgg
     3841 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3901 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3961 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4021 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4081 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4141 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4201 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4261 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4321 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgatg atctcgtcgt
     4381 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4441 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     4501 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4561 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagg
     4621 ggatcaattc tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     4681 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4741 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4801 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4861 tgtatcttaa cgcgtcgagt gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4921 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4981 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5041 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5101 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5161 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5221 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6001 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6061 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6121 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6181 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6241 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6301 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6361 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6421 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6481 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6541 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6601 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6661 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6721 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6781 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6841 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6901 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6961 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
