LOCUS       40924_1264        5448 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector HA-MCS-pcDNA3.1 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5448)
  AUTHORS   Nakashima K, Song SY.
  TITLE     Expression Vector for Mammalian Cell
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5448)
  AUTHORS   Nakashima K, Song SY.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUL-2017) Contact:Kentaro Nakashima Tokushima Bunri 
            University, Institute of Neuroscience; 1314-1 Shido, Sanuki, Kagawa 
            769-2193, Japan
REFERENCE   3  (bases 1 to 5448)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5448)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JUL-2017) Contact:Kentaro Nakashima Tokushima Bunri University, 
            Institute of Neuroscience; 1314-1 Shido, Sanuki, Kagawa 769-2193, 
            Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5448
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             907..933
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_feature    934..1000
                     /label=Multi Cloning Site
                     /note="Multi Cloning Site"
     promoter        complement(1004..1022)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    1048..1272
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1318..1746
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1760..2089
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2156..2947
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3124..3257
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3294..3310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3318..3334)
                     /label=lac operator
                     /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(3342..3372)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3387..3408)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3696..4281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4455..5312)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5313..5417)
                     /label=AmpR promoter
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 gtgtaccgcc
      901 accatgtacc catacgatgt tccagattac gctggatccg ctagcgaatt cgatatcaag
      961 cttggtaccg cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     1021 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1081 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1141 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1201 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     1261 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     1321 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1381 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1441 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1501 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1561 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1621 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1681 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1741 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     1801 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     1861 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     1921 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     1981 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     2041 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     2101 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     2161 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2221 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2281 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2341 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2401 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2461 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2521 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2581 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2641 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     2701 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     2761 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     2821 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     2881 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     2941 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3001 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3061 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3121 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3181 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3241 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3301 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3361 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3421 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3481 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3541 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3601 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3661 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3721 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3781 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3841 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3901 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3961 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4021 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4081 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4141 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4201 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     4261 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     4321 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     4381 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     4441 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     4501 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     4561 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     4621 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     4681 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     4741 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     4801 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     4861 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     4921 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     4981 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     5041 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     5101 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     5161 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     5221 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     5281 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     5341 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     5401 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
