LOCUS       40924_9996        7397 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pcDNA3-N-HA-LIC, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7397)
  AUTHORS   Savitsky PA.
  TITLE     Mammalian expression vector with N-terminal HA tag
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7397)
  AUTHORS   Savitsky PA.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-AUG-2015) Structural Genomics Consortium, University 
            of Oxford, Old Road Campus Research Building, Roosevelt Drive, 
            Oxford, Oxfordshire OX3 7DQ, UK
REFERENCE   3  (bases 1 to 7397)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7397)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-AUG-2015) Structural Genomics Consortium, University of Oxford, 
            Old Road Campus Research Building, Roosevelt Drive, Oxford, 
            Oxfordshire OX3 7DQ, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7397
                     /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             901..927
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     misc_feature    937..950
                     /label=LIC5
                     /note="LIC5"
     promoter        964..1409
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             1410..2828
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     misc_feature    2877..2891
                     /label=LIC3
                     /note="LIC3"
     polyA_signal    2993..3217
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3263..3691
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3705..4035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4102..4893
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5070..5203
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5240..5256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5264..5280)
                     /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(5288..5318)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5333..5354)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5642..6230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6404..7261)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSCGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7262..7366)
                     /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 atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacaccc 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 ggatacccaa gcttatgtcc
      901 tacccatacg acgtaccaga ttacgcggga tccctgtact tccaatccat ggagaccgac
      961 gtccacatat acctgccgtt cactattatt tagtgaaatg agatattatg atattttctg
     1021 aattgtgatt aaaaaggcaa ctttatgccc atgcaacaga aactataaaa aatacagaga
     1081 atgaaaagaa acagatagat tttttagttc tttaggcccg tagtctgcaa atccttttat
     1141 gattttctat caaacaaaag aggaaaatag accagttgca atccaaacga gagtctaata
     1201 gaatgaggtc gaaaagtaaa tcgcgcgggt ttgttactga taaagcaggc aagacctaaa
     1261 atgtgtaaag ggcaaagtgt atactttggc gtcacccctt acatatttta ggtctttttt
     1321 tattgtgcgt aactaacttg ccatcttcaa acaggagggc tggaagaagc agaccgctaa
     1381 cacagtacat aaaaaaggag acatgaacga tgaacatcaa aaagtttgca aaacaagcaa
     1441 cagtattaac ctttactacc gcactgctgg caggaggcgc aactcaagcg tttgcgaaag
     1501 aaacgaacca aaagccatat aaggaaacat acggcatttc ccatattaca cgccatgata
     1561 tgctgcaaat ccctgaacag caaaaaaatg aaaaatataa agttcctgag ttcgattcgt
     1621 ccacaattaa aaatatctct tctgcaaaag gcctggacgt ttgggacagc tggccattac
     1681 aaaacactga cggcactgtc gcaaactatc acggctacca catcgtcttt gcattagccg
     1741 gagatcctaa aaatgcggat gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa
     1801 cttctattga cagctggaaa aacgctggcc gcgtctttaa agacagcgac aaattcgatg
     1861 caaatgattc tatcctaaaa gaccaaacac aagaatggtc aggttcagcc acatttacat
     1921 ctgacggaaa aatccgttta ttctacactg atttctccgg taaacattac ggcaaacaaa
     1981 cactgacaac tgcacaagtt aacgtatcag catcagacag ctctttgaac atcaacggtg
     2041 tagaggatta taaatcaatc tttgacggtg acggaaaaac gtatcaaaat gtacagcagt
     2101 tcatcgatga aggcaactac agctcaggcg acaaccatac gctgagagat cctcactacg
     2161 tagaagataa aggccacaaa tacttagtat ttgaagcaaa cactggaact gaagatggct
     2221 accaaggcga agaatcttta tttaacaaag catactatgg caaaagcaca tcattcttcc
     2281 gtcaagaaag tcaaaaactt ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg
     2341 gcgctctcgg tatgattgag ctaaacgatg attacacact gaaaaaagtg atgaaaccgc
     2401 tgattgcatc taacacagta acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg
     2461 gcaaatggta cctgttcact gactcccgcg gatcaaaaat gacgattgac ggcattacgt
     2521 ctaacgatat ttacatgctt ggttatgttt ctaattcttt aactggccca tacaagccgc
     2581 tgaacaaaac tggccttgtg ttaaaaatgg atcttgatcc taacgatgta acctttactt
     2641 actcacactt cgctgtacct caagcgaaag gaaacaatgt cgtgattaca agctatatga
     2701 caaacagagg attctacgca gacaaacaat caacgtttgc gcctagcttc ctgctgaaca
     2761 tcaaaggcaa gaaaacatct gttgtcaaag acagcatcct tgaacaagga caattaacag
     2821 ttaacaaata aaaacgcaaa agaaaatgcc gatatcctat tggcattgac ggtctccagt
     2881 aaaggtggat acggatctag agcgaagggc gaattctgca gatatccatc acactggcgg
     2941 ccgctcgagc atgcatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3001 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3061 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3121 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3181 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3241 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3301 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3361 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggc
     3421 atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3481 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3541 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3601 tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg gttaaaaaat
     3661 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     3721 gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca tctcaattag
     3781 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3841 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3901 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3961 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4021 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     4081 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4141 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4201 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4261 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4321 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4381 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4441 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4501 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4561 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4621 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4681 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4741 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4801 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4861 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4921 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4981 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5041 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     5101 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     5161 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     5221 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5281 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5341 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5401 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5461 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5521 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5581 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5641 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5701 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5761 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5821 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5881 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5941 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6001 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6061 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     6121 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6181 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6241 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6301 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6361 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6421 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6481 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6541 aagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6601 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6661 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6721 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6781 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6841 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6901 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6961 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7021 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7081 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7141 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7201 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7261 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7321 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7381 aagtgccacc tgacgtc
//
