LOCUS       pBEL2838                6809 bp ds-DNA     circular     15-MAR-2024
DEFINITION  .
KEYWORDS    "Key Features:sfGFP, cytoplasmic" "Backbone:pBEL1764"
            "Construction:Gibson cloning of PCR2919 into EcoRI-NotI-digested
            pBEL1764." "Resistance:Carb" "Location" "Rack" "Box" "Notes"
            "Sequencing information" "Special Category"
COMMENT     Imported using the Genbank importer. File name: pcDNA3.4-TOPO
            (linearized).gb Linearized mammalian vector with 3'-T overhangs and
            bound topoisomerase, for TOPO(R) TA cloning of PCR products and
            high level expression. Do not use phosphorylated primers for the
            PCR amplification because 5' phosphates will prevent ligation into
            this vector.
FEATURES             Location/Qualifiers
     enhancer        47..426
                     /label="CMV enhancer"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          341..360
                     /label="oDCE3730"
                     /note="sequence: ccgcctggcattatgcccag"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     promoter        427..630
                     /label="CMV promoter"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          581..601
                     /label="CMV-Forward"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          637..657
                     /label="oDCE3706"
                     /note="sequence: gtgaaccgtcagatcgcctgg"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          716..805
                     /label="oDCE4021"
                     /note="sequence: ggactctagaggatcgaacccttGGATCTCTAGCGAATTCCCTCTAGACACAGACGCTCACCATGAGCAAAGGAGAAGAACTTTTCACTG"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    739..762
                     /label="Additional 5' UTR from Rb IgH +control plasmid"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    763..777
                     /label="5' UTR Rb IgH mRNA"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    778..780
                     /label="START"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             778..1494
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEFVTAAGITHGMDELYK*"
     misc_feature    781..1491
                     /label="sfGFP"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    1492..1494
                     /label="STOP"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1495..1537
                     /label="Additional 3' UTR from Rb IgH +control plasmid"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    1583..2171
                     /label="WPRE"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(1650..1671)
                     /label="oDCE3707"
                     /note="sequence: aaagcagcgtatccacatagcg"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer          2031..2051
                     /label="oDCE3709"
                     /note="sequence: ggacgtccttctgctacgtcc"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(2209..2228)
                     /label="oDCE3374"
                     /note="sequence: GCGGGTTCCTTCCGGTATTG"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     polyA_signal    2236..2284
                     /label="HSV TK poly(A) signal"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     rep_origin      2486..2914
                     /label="f1 ori"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(2524..2542)
                     /label="oDCE3639"
                     /note="sequence: tcacgctgcgcgtaaccac"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          2546..2566
                     /label="oDCE3710"
                     /note="sequence: ctacacttgccagcgccctag"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        2928..3257
                     /label="SV40 promoter"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          3068..3092
                     /label="oDCE3711"
                     /note="sequence: caattagtcagcaaccatagtcccg"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     rep_origin      3108..3243
                     /label="SV40 ori"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             3324..4118
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          3664..3685
                     /label="oDCE3712"
                     /note="sequence: ccatcatggctgatgcaatgcg"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(3747..3765)
                     /label="oDCE3446"
                     /note="sequence: TCCGAGTACGTGCTCGCTC"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          4267..4287
                     /label="oDCE3713"
                     /note="sequence: ctcatgctggagttcttcgcc"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     polyA_signal    4294..4415
                     /label="SV40 poly(A) signal"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(4331..4350)
                     /label="SV40pA-R"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(4478..4497)
                     /label="M13-48REV"
                     /note="sequence: CGGATAACAATTTCACACAG"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          4858..4879
                     /label="oDCE3449"
                     /note="sequence: CTGGCGTTTTTCCATAGGCTCC"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     rep_origin      complement(4866..5454)
                     /label="ori"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(5063..5084)
                     /label="oDCE3562"
                     /note="sequence: ctgagatacctacagcgtgagc"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(5170..5188)
                     /label="oDCE3451"
                     /note="sequence: CCGGGTTGGACTCAAGACG"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          5474..5493
                     /label="oDCE3548"
                     /note="sequence: ctacggggtctgacgctcag"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             complement(5625..6485)
                     /label="AmpR"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(5678..5699)
                     /label="oDCE3561"
                     /note="sequence: gggagtcaggcaactatggatg"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          6077..6096
                     /label="oDCE3547"
                     /note="sequence: gaagtaagttggccgcagtg"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(6298..6318)
                     /label="oDCE3560"
                     /note="sequence: gagttttcgccccgaagaacg"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     promoter        complement(6486..6590)
                     /label="AmpR promoter"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          6674..6696
                     /label="oDCE3714"
                     /note="sequence: ctctgatgccgcatagttaagcc"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
ORIGIN
        1 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
       61 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      121 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      601 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat cgcctggaga
      661 cgccatccac gctgttttga cctccataga agacaccggg accgatccag cctccggact
      721 ctagaggatc gaacccttGG ATCTCTAGCG AATTCCCTCT AGACACAGAC GCTCACCATG
      781 AGCAAAGGAG AAGAACTTTT CACTGGAGTT GTCCCAATTC TTGTTGAATT AGATGGTGAT
      841 GTTAATGGGC ACAAATTTTC TGTCCGTGGA GAGGGTGAAG GTGATGCTAC AAACGGAAAA
      901 CTCACCCTTA AATTTATTTG CACTACTGGA AAACTACCTG TTCCGTGGCC AACACTTGTC
      961 ACTACTCTGA CCTATGGTGT TCAATGCTTT TCCCGTTATC CGGATCACAT GAAACGGCAT
     1021 GACTTTTTCA AGAGTGCCAT GCCCGAAGGT TATGTACAGG AACGCACTAT ATCTTTCAAA
     1081 GATGACGGGA CCTACAAGAC GCGTGCTGAA GTCAAGTTTG AAGGTGATAC CCTTGTTAAT
     1141 CGTATCGAGT TAAAGGGTAT TGATTTTAAA GAAGATGGAA ACATTCTTGG ACACAAACTC
     1201 GAGTACAACT TTAACTCACA CAATGTATAC ATCACGGCAG ACAAACAAAA GAATGGAATC
     1261 AAAGCTAACT TCAAAATTCG CCACAACGTT GAAGATGGTT CCGTTCAACT AGCAGACCAT
     1321 TATCAACAAA ATACTCCAAT TGGCGATGGC CCTGTCCTTT TACCAGACAA CCATTACCTG
     1381 TCGACACAAT CTGTCCTTTC GAAAGATCCC AACGAAAAGC GTGACCACAT GGTCCTTCTT
     1441 GAGTTTGTAA CTGCTGCTGG GATTACACAT GGCATGGATG AGCTCTACAA ATGAGCGGCC
     1501 GCTCGAGGCC GGCAAGGCCG GATCCCCCGA CCTCGACaag ggttcgatcc ctaccggtta
     1561 gtaatgagtt tgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac
     1621 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     1681 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     1741 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     1801 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     1861 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     1921 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     1981 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     2041 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     2101 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     2161 cgcctccccg cctggaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     2221 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt
     2281 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     2341 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     2401 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc
     2461 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2521 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2581 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2641 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2701 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2761 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2821 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2881 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     2941 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3001 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3061 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     3121 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     3181 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     3241 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
     3301 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     3361 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     3421 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     3481 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     3541 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     3601 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     3661 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     3721 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     3781 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     3841 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     3901 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     3961 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4021 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4081 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa
     4141 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4201 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4261 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4321 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4381 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4441 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4501 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4561 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4621 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4681 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     4741 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4801 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4861 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4921 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4981 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5041 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5101 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5161 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5221 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5281 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5341 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5401 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5461 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5521 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5581 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5641 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     5701 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     5761 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5821 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5881 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5941 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6001 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6061 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6121 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6181 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6241 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6301 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6361 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6421 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6481 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6541 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6601 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     6661 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     6721 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     6781 acaattgcat gaagaatctg cttagggtt
//