LOCUS       pcDNA_ZEO_DD_Nluc_QtSig_corrected        5907 bp    DNA     circular UNA 01-OCT-2024
DEFINITION  .
ACCESSION   urn.local...v-ixztrrm
VERSION     urn.local...v-ixztrrm
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     940..983
                     /Tm=75
                     /Sequence="TCCAGTGTGGTGGAATTCGCCACCATGGGAGTGCAGGTGGAAAC"
                     /Description="<html><body></body></html>"
                     /label="DD-Eos_GeoSig_GA_for"
     primer_bind     complement(941..963)
                     /Tm=65
                     /Sequence="GGTGGCGAATTCCACCACACTGG"
                     /Description="<html><body></body></html>"
                     /label="pcDNA_Zeo_lin_rev"
     primer_bind     943..963
                     /Tm=62
                     /Sequence="AGTGTGGTGGAATTCGCCACC"
                     /Description="<html><body></body></html>"
                     /label="DD-Eos_OL_for"
     misc_feature    958..963
                     /label="Kozak"
     CDS             join(964..966,967..1287)
                     /codon_start=1
                     /note="L106P mutant of FKBP12"
                     /product="destabilization domain that can be stabilized by
                     Shield1 in the ProteoTuner™ system"
                     /transl_table=1
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKVDSSRDRN
                     KPFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDV
                     ELLKPE"
                     /label="DD-N"
     primer_bind     complement(1252..1266)
                     /Tm=49
                     /Sequence="ATCGAAGACGAGAGT"
                     /Extension="GGAGGAGGA"
                     /Description="<html><body></body></html>"
                     /label="Primer 1"
     primer_bind     1256..1320
                     /Tm=75
                     /Sequence="TCGTCTTCGATGTGGAGCTTCTAAAACCGGAAGGATCTGGGGTGTTC
                     ACCCTGGAAGATTTCGTG"
                     /Description="<html><body></body></html>"
                     /label="DD-N_NLuc_GA_for"
     primer_bind     complement(1265..1295)
                     /Tm=65
                     /Sequence="CCAGATCCTTCCGGTTTTAGAAGCTCCACAT"
                     /Description="<html><body></body></html>"
                     /label="pcDNA_ZEO_lin_rev(neu)"
     CDS             1288..1296
                     /codon_start=1
                     /transl_table=1
                     /translation="GSG"
                     /label="Linker"
     CDS             1297..1806
                     /codon_start=1
                     /transl_table=1
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDG
                     VTPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVT
                     GWRLCERILA"
                     /label="Nluc"
     primer_bind     complement(1788..1846)
                     /Tm=81
                     /Sequence="TGAAGCCCTTAGATCCGCCTCCGCCAGATCCTCCGCCGCCGGCCAGG
                     ATTCTCTCGCAC"
                     /Description="<html><body></body></html>"
                     /label="NLuc_Linker_QtSig_GA_rev"
     CDS             1807..1836
                     /codon_start=1
                     /transl_table=1
                     /translation="GGGGSGGGGS"
                     /label="Linker"
     primer_bind     1807..1826
                     /Tm=68
                     /Sequence="GGCGGCGGAGGATCTGGCGG"
                     /Description="<html><body></body></html>"
                     /label="pcDNA_ZEO_lin_for(neu)"
     primer_bind     complement(1816..1886)
                     /Tm=81
                     /Sequence="TAGACTCGAGCGGCCGCTCATTGGATCAGGCTGCCCACGGTGAAGCC
                     CTTAGATCCGCCTCCGCCAGATCC"
                     /Description="<html><body></body></html>"
                     /label="DD-mEos4b_GeoTag_rev"
     CDS             1837..1866
                     /codon_start=1
                     /transl_table=1
                     /translation="KGFTVGSLIQ"
                     /label="GeoTag"
     primer_bind     complement(1848..1877)
                     /Tm=55
                     /Sequence="GCGCCAGAGCCTTGGATCAGGCTGCCCACG"
                     /Mismatch_Positions="===C=A=AG=C==================="
                     /Mismatches=5
                     /Extension="GCTGAAGTTTGTG"
                     /Description="<html><body></body></html>"
                     /label="DD-Eos_GeoSig_GA_rev"
     primer_bind     complement(1851..1869)
                     /Tm=54
                     /Sequence="TCATTGGATCAGGCTGCCC"
                     /Extension="GCTCCCTTAGCTCCCTGAGACG"
                     /Description="<html><body></body></html>"
                     /label="mEos4b_BM40_GA_rev"
     primer_bind     1870..1889
                     /Tm=63
                     /Sequence="GCGGCCGCTCGAGTCTAGAG"
                     /Description="<html><body></body></html>"
                     /label="pcDNA_Zeo_lin_for"
     polyA_signal    1920..2144
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2190..2618
                     /snapgene_direction="RIGHT"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2632..2962
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     rep_origin      2813..2948
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     promoter        3010..3057
                     /note="synthetic bacterial promoter&nbsp;"
                     /label="EM7 promoter"
     CDS             3076..3450
                     /codon_start=1
                     /gene="<i>Sh&nbsp;ble</i> from&nbsp;<i>Streptoalloteichus
                     hindustanus</i>"
                     /note="confers resistance to bleomycin, phleomycin, and
                     Zeocin™"
                     /product="antibiotic-binding protein"
                     /transl_table=1
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRD
                     DVTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPW
                     GREFALRDPAGNCVHFVAEEQD*"
                     /label="BleoR"
     polyA_signal    3580..3701
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(3750..3766)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    3774..3790
                     /bound_moiety="<i>lac</i> repressor encoded by
                     <i>lacI</i>"
                     /note="The <i>lac</i> repressor binds to the <i>lac</i>
                     operator to inhibit transcription in <i>E. coli</i>. This
                     inhibition can be relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        join(complement(3823..3828),complement(3805..3822),
                     complement(3798..3804))
                     /note="promoter for the <i>E. coli lac</i> operon"
                     /label="lac promoter"
     protein_bind    3843..3864
                     /bound_moiety="<i>E. coli</i> catabolite activator
                     protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4152..4740)
                     /snapgene_direction="LEFT"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             join(complement(5703..5771),complement(4911..5702))
                     /codon_start=1
                     /gene="<i>bla</i>"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
                     /label="AmpR"
     promoter        complement(5772..5876)
                     /gene="<i>bla</i>"
                     /label="AmpR promoter"
ORIGIN      
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatgggag tgcaggtgga aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc
     1021 ggccagacct gtgtggtgca ctacaccggg atgcttgaag atggaaagaa agtcgattcc
     1081 tcccgggaca gaaacaagcc ctttaagttt atgctaggca agcaggaggt gatccgaggc
     1141 tgggaagaag gggttgccca gatgagtgtg ggtcagagag ccaaactgac tatatctcca
     1201 gattatgcct atggtgccac tgggcaccca ggcatcatcc caccacatgc cactctcgtc
     1261 ttcgatgtgg agcttctaaa accggaagga tctggggtgt tcaccctgga agatttcgtg
     1321 ggcgactggc ggcagaccgc tggctacaat ctggaccagg tgctggaaca gggcggcgtg
     1381 tcctctctgt ttcagaacct gggcgtgtcc gtgaccccca tccagagaat cgtgctgagc
     1441 ggcgagaacg gcctgaagat cgacatccac gtgatcatcc cttacgaggg cctgtccggc
     1501 gaccagatgg gccagatcga gaagatcttt aaggtggtgt accccgtgga cgaccaccac
     1561 ttcaaagtga tcctgcacta cggcaccctc gtgatcgacg gcgtgacccc taacatgatc
     1621 gactacttcg gcagacccta cgagggaatc gccgtgttcg acggcaagaa aatcaccgtg
     1681 accggcaccc tgtggaacgg caacaagatc atcgacgaga gactgatcaa ccccgacggc
     1741 agcctgctgt tcagagtgac catcaatggc gtgacaggct ggcggctgtg cgagagaatc
     1801 ctggccggcg gcggaggatc tggcggaggc ggatctaagg gcttcaccgt gggcagcctg
     1861 atccaatgag cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     1921 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1981 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2041 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2101 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2161 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2221 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2281 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2341 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2401 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2461 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2521 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     2581 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2641 ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct
     2701 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2761 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     2821 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     2881 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     2941 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3001 tcagcacgtg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3061 gtgaggaact aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac
     3121 gtcgccggag cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag
     3181 gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac
     3241 caggtggtgc cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac
     3301 gccgagtggt cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc
     3361 gagatcggcg agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc
     3421 gtgcacttcg tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc
     3481 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3541 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     3601 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3661 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     3721 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     3781 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     3841 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     3901 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3961 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4021 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4081 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4141 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4201 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4261 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4321 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     4381 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4441 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4501 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4561 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4621 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4681 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4741 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4801 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4861 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4921 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4981 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5041 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5101 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5161 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5221 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5281 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5341 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5401 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5461 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5521 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5581 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5641 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5701 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5761 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5821 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5881 tttccccgaa aagtgccacc tgacgtc
//
