LOCUS       pcDNA-c/o_NL4-3_Env_delta_CT        7533 bp ds-DNA     circular     20-JAN-2020
DEFINITION  NL4-3 Env lacking cytoplasmic tail expression vector.
ACCESSION   urn.local...18-actak2j
KEYWORDS    .
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
            .
COMMENT     ApEinfo:methylated:1.
FEATURES             Location/Qualifiers
     promoter        236..852
                     /ApEinfo_fwdcolor="#ffff80"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="CMV_immearly_promoter"
     misc_feature    588..593
                     /Recognition_pattern="TAC^GTA"
                     /annotation_group="TACGTA"
                     /label="SnaBI"
                     /note="Geneious type: restriction site"
     promoter        863..881
                     /ApEinfo_fwdcolor="#ffff80"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="T7_promoter"
     misc_feature    915..920
                     /Recognition_pattern="T^CTAGA"
                     /annotation_group="TCTAGA"
                     /label="XbaI"
                     /note="Geneious type: restriction site"
     CDS             927..3059
                     /created_by="wdiehl"
                     /label="codon optimized NL4-3 Env delta CT"
     ORF             927..3059
                     /Genetic_code="Standard"
                     /Reading_frame=3
                     /created_by="ORF Finder"
                     /label="ORF 5 (frame 3)"
     misc_feature    1031..1037
                     /Recognition_pattern="G^GTNACC"
                     /annotation_group="GGTNACC"
                     /label="BstEII"
                     /note="Geneious type: restriction site"
     misc_feature    1806..1811
                     /Recognition_pattern="T^GTACA"
                     /annotation_group="TGTACA"
                     /label="BsrGI"
                     /note="Geneious type: restriction site"
     misc_feature    1853..1859
                     /Recognition_pattern="RG^GWCCY"
                     /annotation_group="RGGWCCY"
                     /label="PpuMI"
                     /note="Geneious type: restriction site"
     misc_feature    1919..1924
                     /Recognition_pattern="C^TCGAG"
                     /annotation_group="CTCGAG"
                     /label="XhoI"
                     /note="Geneious type: restriction site"
     misc_feature    3060..3065
                     /Recognition_pattern="G^AATTC"
                     /annotation_group="GAATTC"
                     /label="EcoRI"
                     /note="Geneious type: restriction site"
     source          3066..914
                     /organism="pcDNA3.1(-)"
                     /mol_type="other DNA"
                     /ApEinfo_label="source:pcDNA3.1(-)"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /modified_by="wdiehl"
                     /label="source:pcDNA3.1(-)"
     misc_feature    3095..3100
                     /Recognition_pattern="GGTAC^C"
                     /annotation_group="GGTACC"
                     /label="KpnI"
                     /note="Geneious type: restriction site"
     misc_feature    3101..3106
                     /Recognition_pattern="A^AGCTT"
                     /annotation_group="AAGCTT"
                     /label="HindIII"
                     /note="Geneious type: restriction site"
     terminator      3130..3357
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="bGH_PA_terminator"
     rep_origin      3420..3726
                     /ApEinfo_fwdcolor="#ffcaff"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="f1_origin"
     promoter        3858..4126
                     /ApEinfo_fwdcolor="#ffff80"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="SV40_promoter"
     rep_origin      4025..4102
                     /ApEinfo_fwdcolor="#ffcaff"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="SV40_origin"
     misc_feature    4159..4164
                     /Recognition_pattern="C^CTAGG"
                     /annotation_group="CCTAGG"
                     /label="AvrII"
                     /note="Geneious type: restriction site"
     gene            4244..5032
                     /gene="NeoR/KanR"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="NeoR/KanR"
     misc_feature    4275..4280
                     /Recognition_pattern="C^GGCCG"
                     /annotation_group="CGGCCG"
                     /label="EagI"
                     /note="Geneious type: restriction site"
     terminator      5212..5331
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
                     /ApEinfo_fwdcolor="pink"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="SV40_PA_terminator"
     promoter        complement(5375..5393)
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
                     /ApEinfo_fwdcolor="#ffff80"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="M13_reverse_primer"
     promoter        complement(5428..5457)
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
                     /ApEinfo_fwdcolor="#ffff80"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="lac_promoter"
     gene            complement(6537..7397)
                     /gene="Ampicillin"
                     /translation="MAGWASLGRSFRTPESRSEELVKKAIEGDALRIGSGDTVKHEEA
                     VSPFAAKLFSNITGSQRYVLIAVRHTQPATVDESRKAAIFHHDIRQAGIAMGHDEILA
                     VGHARLEPGEQFGWREPLMLFVQIILIDKTGFHPSTCSLDAMFRLVVEWAGSRIKRMQ
                     PPHCISHDGYFLGRSKVR*"
                     /ApEinfo_fwdcolor="#408080"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /label="Ampicillin"
     promoter        complement(7439..7467)
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /ApEinfo_fwdcolor="#ffff80"
                     /ApEinfo_revcolor="pink"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0"
                     /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 gctggctagc
      901 gtttaaacgg gccctctaga gccgccatga gagtgaagga aaaatatcag cacctctgga
      961 gatgggggtg gaagtgggga acgatgctcc tcgggattct gatgatctgt agcgccactg
     1021 aaaagctctg ggtcaccgtt tactatggtg tccctgtgtg gaaggaggct acaaccacac
     1081 tgttctgtgc cagcgatgca aaggcatatg acacagaggt tcataatgtc tgggcaaccc
     1141 acgcctgcgt tccgaccgat ccgaatcccc aagaagttgt cctcgtcaac gtcacagaga
     1201 atttcaatat gtggaaaaat gacatggttg agcagatgca cgaagacata attagcctgt
     1261 gggatcaatc cctgaaacca tgcgtgaagc tcacgccgct ctgcgtgtct ctcaagtgca
     1321 ccgaccttaa gaatgatacg aatacaaact ctagtagcgg acggatgatt atggaaaaag
     1381 gcgaaatcaa gaactgctcc ttcaatatat caacaagcat ccgagataaa gtccagaagg
     1441 aatacgcatt tttttataaa ttggatattg tccccataga taataccagt tatcgactta
     1501 tttcttgcaa tacgagcgta atcacacaag cgtgcccgaa ggtttccttc gaacctatac
     1561 ctattcatta ttgcgcacca gcaggatttg cgatcttgaa atgcaacaat aagacgttta
     1621 atgggacagg cccatgcacc aacgtatcta cggtccagtg cactcatggt atcagacctg
     1681 ttgtctcaac tcaactgttg ctcaacggct ctctggcaga agaggatgtc gtcatccgat
     1741 ctgccaattt tacggataac gctaagacaa ttattgtgca actgaacacg tctgtcgaga
     1801 taaattgtac aagaccaaat aacaatacgc ggaagagcat ccgcatccaa cgaggtcccg
     1861 ggagagcttt tgttacgatc ggcaagatag ggaatatgag acaggcgcac tgtaatatct
     1921 cgagggctaa gtggaacgca acgcttaaac aaatagctag caagttgagg gagcagttcg
     1981 gaaacaataa gacaattatc tttaaacaat cctcaggagg ggatccagaa atcgtgacac
     2041 actctttcaa ctgcgggggc gagttttttt attgtaattc cactcagttg ttcaatagca
     2101 cgtggtttaa ctcaacgtgg tcaactgagg gttccaacaa taccgaaggc tctgatacta
     2161 ttaccctgcc ttgccggata aagcagttta ttaacatgtg gcaggaagtt ggcaaagcta
     2221 tgtatgcacc cccaatctcc ggacagataa ggtgctcatc caatataact ggacttctgc
     2281 tgacacggga tggcgggaat aacaacaacg gaagcgaaat ttttcgacct ggtgggggtg
     2341 atatgcgaga caattggcgg agcgagctct acaagtacaa agtcgttaag atcgagcccc
     2401 tcggagttgc gcctaccaaa gctaaacgac gagtggtaca aagggagaag cgagcggtag
     2461 gcataggcgc gctcttcttg ggtttccttg gtgctgcagg gtctactatg ggcgcagcct
     2521 ctatgacact cactgtccag gctcggcagc tgctgtccga catagtccag caacagaaca
     2581 accttctccg cgccattgaa gctcagcaac acctccttca gctcacggtc tggggaataa
     2641 aacagctcca ggcacgcata ttggcagtgg agaggtatct taaggatcaa caactgctgg
     2701 gtatatgggg gtgtagtgga aagttgatct gcacaacagc agtcccctgg aacgcctctt
     2761 ggagcaataa gtctcttgaa caaatctgga ataatatgac ctggatggag tgggaccgcg
     2821 agattaataa ttacacgagt ctgattcata gtcttattga agagtcacag aatcagcaag
     2881 agaagaacga acaggagttg ctcgaattgg ataagtgggc ttccctctgg aactggttta
     2941 acatcactaa ctggctgtgg tacataaagt tgttcatcat gatcgtaggg ggcctcgtag
     3001 gcctccgcat agtgtttgcc gtgctcagta tagtgaatcg cgtaagacaa ggttattaag
     3061 aattccacca cactggacta gtggatccga gctcggtacc aagcttaagt ttaaaccgct
     3121 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3181 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3241 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3301 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3361 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3421 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3481 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3541 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3601 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3661 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3721 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3781 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3841 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3901 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3961 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4021 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4081 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4141 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4201 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4261 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4321 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4381 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4441 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4501 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4561 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4621 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4681 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4741 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4801 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4861 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4921 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4981 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5041 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     5101 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5161 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5221 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5281 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5341 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5401 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5461 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5521 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5581 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5641 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5701 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5761 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5821 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5881 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5941 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6001 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6061 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6121 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6181 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6241 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6301 accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     6361 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     6421 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     6481 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     6541 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6601 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6661 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6721 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6781 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6841 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6901 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6961 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     7021 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     7081 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     7141 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     7201 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     7261 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     7321 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     7381 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     7441 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7501 cgcacatttc cccgaaaagt gccacctgac gtc
//
