LOCUS hUBC_hsPUM1_iRFP670 6060 bp DNA circular UNA 20-APR-2018 DEFINITION Concatenation of 2 sequences (Circularized). ACCESSION urn.local...29-8x94ucb VERSION urn.local...29-8x94ucb KEYWORDS . SOURCE ORGANISM . FEATURES Location/Qualifiers CDS complement(1..861) /gene="bla" /note="ampR (confers resistance to ampicillin)" /codon_start=1 /product="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" /label="bla CDS" gene complement(1..861) /gene="bla" /label="bla gene" misc_feature 552..557 /Recognition_pattern="AGT^ACT" /annotation_group="AGTACT" /label="ScaI, AssI, BmcAI, ZrmI" /note="Geneious type: restriction site" misc_feature 590..601 /Recognition_pattern="(10/12)CGANNNNNNTGC(12/10)" /annotation_group="CGANNNNNNTGC" /label="BcgI" /note="Geneious type: restriction site" misc_feature 669..678 /Recognition_pattern="GAANN^NNTTC" /annotation_group="GAANNNNTTC" /label="XmnI, Asp700I, MroXI, PdmI" /note="Geneious type: restriction site" sig_peptide complement(793..861) /gene="bla" /note="required for secretion to the periplasm; cleaved off to form the mature beta-lactamase protein." /label="bla signal peptide" misc_feature 876..881 /Recognition_pattern="AAT^ATT" /annotation_group="AATATT" /label="SspI" /note="Geneious type: restriction site" -10_signal complement(905..910) /note="bla promoter (counter-clockwise); GAGACA; transcript start 2548 (complementary strand)" /label="-10 signal" -35_signal complement(926..931) /note="bla promoter (counter-clockwise); TTCAAA" /label="-35 signal" misc_feature 1050^1051 /Original_Bases="GCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACC TCTGACACATGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAG CAGACAAGCCCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTTAA" /label="GCCCTTTCGTCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGACACA TGCAGCTCCCGGAGACGGTCACAGCTTGTCTGTAAGCGGATGCCGGGAGCAGACAAGC CCGTCAGGGCGCGTCAGCGGGTGTTGGCGGGTGTCGGGGCTGGCTTAA" /note="Geneious type: Editing History Deletion" source order(<1051..>1316,<4856..>1050) /organism="Cloning vector pNEB193" /mol_type="other DNA" /label="source Cloning vector pNEB193" misc_feature 1084..1094 /Recognition_pattern="GCANNNN^NTGC" /annotation_group="GCANNNNNTGC" /label="BstAPI" /note="Geneious type: restriction site" misc_feature 1088..1093 /Recognition_pattern="CA^TATG" /annotation_group="CATATG" /label="NdeI, FauNDI" /note="Geneious type: restriction site" primer_bind 1257..1280 /created_by="User" /label="M13F (-47) Primer from GeneWiz" primer_bind 1283..1300 /created_by="User" /label="M13F(-21) Primer From GeneWiz" misc_feature 1301..1306 /Recognition_pattern="G^AATTC" /annotation_group="GAATTC" /label="EcoRI" /note="Geneious type: restriction site" misc_feature 1307..1312 /Recognition_pattern="GAG^CTC" /annotation_group="GAGCTC" /label="Ecl136II, Eco53kI, EcoICRI" /note="Geneious type: restriction site" misc_feature 1307..1312 /Recognition_pattern="GAGCT^C" /annotation_group="GAGCTC" /label="SacI, Psp124BI, SstI" /note="Geneious type: restriction site" misc_feature 1331..1335 /Original_Bases /label /note="Geneious type: Editing History Insertion" misc_feature 1336..2539 /created_by="User" /modified_by="User" /label="human_UBC_promoter" misc_feature 1356..1361 /Recognition_pattern="CCGC^GG" /annotation_group="CCGCGG" /label="SacII, Cfr42I, KspI, Sfr303I, SgrBI, SstII" /note="Geneious type: restriction site" misc_feature 1382..1387 /Recognition_pattern="AGC^GCT" /annotation_group="AGCGCT" /label="Eco47III, AfeI, Aor51HI" /note="Geneious type: restriction site" misc_feature 1411 /Original_Bases /label /note="Geneious type: Editing History Insertion" misc_feature 1417^1418 /Original_Bases="T" /label="T" /note="Geneious type: Editing History Deletion" misc_feature complement(1438..1444) /Recognition_pattern="CCTNAGC(-5/-2)" /annotation_group="CCTNAGC" /label="Bpu10I" /note="Geneious type: restriction site" misc_feature 1480..1490 /Recognition_pattern="(10/15)ACNNNNGTAYC(12/7)" /annotation_group="ACNNNNGTAYC" /label="BaeI" /note="Geneious type: restriction site" misc_feature 2559..2567 /created_by="User" /label="Kozak_Sequence" misc_feature 2564..2590 /created_by="User" /label="NLS" misc_feature 2597..2623 /created_by="User" /label="HA_TAG" misc_feature 2630..3676 /created_by="User" /label="hsPUM1" misc_feature 3683..3706 /ApEinfo_fwdcolor="#ffff80" /ApEinfo_revcolor="#ffff80" /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0" /label="10 aa linker" misc_feature 3707..3712 /created_by="User" /label="BspEI site" misc_feature 3707..3712 /Recognition_pattern="T^CCGGA" /annotation_group="TCCGGA" /label="AccIII, Aor13HI, BseAI, Bsp13I, BspEI, Kpn2I, MroI" /note="Geneious type: restriction site" misc_feature 3709 /Original_Bases="G" /label="G" /note="Geneious type: Editing History Replacement" ORF 3713..4729 /created_by="User" /modified_by="User" /label="iRFP670" misc_feature 4730..4735 /created_by="User" /label="SalI Site" terminator 4736..4855 /label="SV40_PA_terminator" misc_feature 4735^4736 /Original_Bases="GGAGACGGAATTCCGTCTCA" /label="GGAGACGGAATTCCGTCTCA" /note="Geneious type: Editing History Deletion" source <4736..>4855 /organism="pcDNA3.1(+)" /mol_type="other DNA" /label="source pcDNA3.1(+)" misc_feature complement(4770..4789) /created_by="User" /modified_by="User" /label="SV40pA Rev Primer from Genewiz" /note="Geneious type: primer_bind_reverse" misc_feature complement(4813..4818) /Recognition_pattern="GAATGC(1/-1)" /annotation_group="GAATGC" /label="BsmI, BsaMI, Mva1269I, PctI" /note="Geneious type: restriction site" primer_bind 4824..4843 /modified_by="User" /label="EBV_rev_primer" misc_feature 4862..4869 /Recognition_pattern="GTTT^AAAC" /annotation_group="GTTTAAAC" /label="PmeI, MssI" /note="Geneious type: restriction site" misc_feature 4876..4881 /Recognition_pattern="GCATG^C" /annotation_group="GCATGC" /label="SphI, BbuI, PaeI" /note="Geneious type: restriction site" misc_feature complement(4900..4916) /created_by="User" /label="M13R Primer From GeneWiz" /note="Geneious type: primer_bind_reverse" -10_signal complement(4949..4954) /note="Plac promoter (drives lacZalpha; counter-clockwise); TATGTT" /label="-10 signal" -35_signal complement(4973..4978) /note="Plac promoter (drives lacZalpha; counter-clockwise); TTTACA" /label="-35 signal" protein_bind complement(4998..5010) /note="CAP protein binding site" /bound_moiety="catabolite activator protein (CAP)" /label="CAP protein binding site protein bind" misc_feature 5118..5124 /Recognition_pattern="GCTCTTC(1/4)" /annotation_group="GCTCTTC" /label="SapI, BspQI, LguI, PciSI" /note="Geneious type: restriction site" misc_feature 5241..5246 /Recognition_pattern="A^CATGT" /annotation_group="ACATGT" /label="PciI, PscI" /note="Geneious type: restriction site" rep_origin complement(5302..5890) /note="pMB1 origin of replication (counter-clockwise) (RNAII -35 to RNA/DNA switch point)" /label="rep origin" misc_RNA complement(5302..5854) /note="RNAII transcript (complementary strand)" /label="RNA" -35_signal 5708..5713 /note="RNAI promoter (clockwise); TTGAAG" /label="RNAI promoter (clockwise); TTGAAG -35 signal" -10_signal 5730..5735 /note="RNAI promoter (clockwise); GCTACA" /label="RNAI promoter (clockwise); GCTACA -10 signal" misc_RNA 5744..5851 /note="RNAI transcript" /label="RNAI transcript" -10_signal complement(5864..5869) /note="RNAII promoter (counter-clockwise); CGTAAT" /label="-10 signal" -35_signal complement(5885..5890) /note="RNAII promoter (counter-clockwise); TTGAGA" /label="-35 signal" ORIGIN 1 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat 61 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc 121 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa 181 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca 241 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa 301 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt 361 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc 421 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact 481 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc 541 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg 601 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct 661 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc 721 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag 781 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac 841 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg 901 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt 961 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac 1021 attaacctat aaaaataggc gtatcacgag ctatgcggca tcagagcaga ttgtactgag 1081 agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag 1141 gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc 1201 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc 1261 agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc tcggtacacg 1321 tatgtcgagg taggcgcgcc gggttttggc gcctcccgcg ggcgcccccc tcctcacggc 1381 gagcgctgcc acgtcagacg aagggcgcag cgagcgtcct gatccttccg cccggacgct 1441 caggacagcg gcccgctgct cataagactc ggccttagaa ccccagtatc agcagaagga 1501 cattttagga cgggacttgg gtgactctag ggcactggtt ttctttccag agagcggaac 1561 aggcgaggaa aagtagtccc ttctcggcga ttctgcggag ggatctccgt ggggcggtga 1621 acgccgatga ttatataagg acgcgccggg tgtggcacag ctagttccgt cgcagccggg 1681 atttgggtcg cggttcttgt ttgtggatcg ctgtgatcgt cacttggtga gtagcgggct 1741 gctgggctgg ccggggcttt cgtggccgcc gggccgctcg gtgggacgga agcgtgtgga 1801 gagaccgcca agggctgtag tctgggtccg cgagcaaggt tgccctgaac tgggggttgg 1861 ggggagcgca gcaaaatggc ggctgttccc gagtcttgaa tggaagacgc ttgtgaggcg 1921 ggctgtgagg tcgttgaaac aaggtggggg gcatggtggg cggcaagaac ccaaggtctt 1981 gaggccttcg ctaatgcggg aaagctctta ttcgggtgag atgggctggg gcaccatctg 2041 gggaccctga cgtgaagttt gtcactgact ggagaactcg gtttgtcgtc tgttgcgggg 2101 gcggcagtta tggcggtgcc gttgggcagt gcacccgtac ctttgggagc gcgcgccctc 2161 gtcgtgtcgt gacgtcaccc gttctgttgg cttataatgc agggtggggc cacctgccgg 2221 taggtgtgcg gtaggctttt ctccgtcgca ggacgcaggg ttcgggccta gggtaggctc 2281 tcctgaatcg acaggcgccg gacctctggt gaggggaggg ataagtgagg cgtcagtttc 2341 tttggtcggt tttatgtacc tatcttctta agtagctgaa gctccggttt tgaactatgc 2401 gctcggggtt ggcgagtgtg ttttgtgaag ttttttaggc accttttgaa atgtaatcat 2461 ttgggtcaat atgtaatttt cagtgttaga ctagtaaatt gtccgctaaa ttctggccgt 2521 ttttggcttt tttgttagac ctggaggtac cgccaccgcc accatggctc caaagaagaa 2581 gcgtaaggta ggcagctacc catacgatgt tccagattac gctggcagcg gtcggtcccg 2641 gctcttggag gactttcgaa acaatcgata tcctaacttg cagcttcgcg agatcgctgg 2701 tcatattatg gaattctctc aggatcaaca cgggtctaga ttcattcagc tcaagctgga 2761 gagagccaca cctgccgaac ggcagctcgt gtttaatgag atcctccagg ctgcctatca 2821 actgatggtg gacgtgttcg gcaattacgt catccagaag ttctttgaat ttggtagtct 2881 cgagcagaag ctggcccttg cagagagaat ccgcggccat gtactgtccc ttgccctgca 2941 gatgtacggg tgccgcgtaa tccagaaggc actggaattt ataccgtccg accagcagaa 3001 tgagatggtg cgggaactcg atggtcatgt gttgaaatgc gtaaaagatc agaacggtaa 3061 ccacgtggtt cagaaatgta tcgagtgtgt ccagccacag agcttgcaat tcatcataga 3121 tgcctttaag ggccaggttt tcgctctgag cacgcacccc tacggatgcc gggtcattca 3181 gcggattctt gaacattgtc tgccagacca gactctgcct atcctggagg agctccacca 3241 acacaccgaa cagttggtcc aggatcaata tggtaattac gttattcagc atgtgctgga 3301 acatgggcgg cctgaggaca agtccaaaat tgtagctgag attcgaggca atgtgctcgt 3361 tctttcccag cacaagttcg cttctaacgt cgtggagaaa tgtgtgactc acgccagccg 3421 gactgagcgc gcggtgctca ttgatgaagt gtgtaccatg aacgatgggc cacatagcgc 3481 actttacaca atgatgaaag accagtacgc aaactacgtg gttcagaaga tgatagacgt 3541 cgctgaaccg gggcagcgca aaattgtcat gcacaagatc cgcccacaca tagctactct 3601 tcggaaatac acctatggca agcacatcct cgcaaagctg gagaagtatt acatgaaaaa 3661 cggggtggat ctgggaggat ccaagggctc gggctcgacc tcgggctccg gaatgtcgcg 3721 gaggcggcac agctacgaga acgatggtgg acaacctcac aaaaggagga agacgtctcc 3781 accggtcgcc accatggcgc gtaaggtcga tctcacctcc tgcgatcgcg agccgatcca 3841 catccccggc agcattcagc cgtgcggctg cctgctagcc tgcgacgcgc aggcggtgcg 3901 gatcacgcgc attacggaaa atgccggcgc gttctttgga cgcgaaactc cgcgggtcgg 3961 tgagctactc gccgattact tcggcgagac cgaagcccat gcgctgcgca acgcactggc 4021 gcagtcctcc gatccaaagc gaccggcgct gatcttcggt tggcgcgacg gcctgaccgg 4081 ccgcaccttc gacatctcac tgcatcgcca tgacggtaca tcgatcatcg agttcgagcc 4141 tgcggcggcc gaacaggccg acaatccgct gcggctgacg cggcagatca tcgcgcgcac 4201 caaagaactg aagtcgctcg aagagatggc cgcacgggtg ccgcgctatc tgcaggcgat 4261 gctcggctat caccgcgtga tgttgtaccg cttcgcggac gacggctccg ggatggtgat 4321 cggcgaggcg aagcgcagcg acctcgagag ctttctcggt cagcactttc cggcgtcgct 4381 ggtcccgcag caggcgcggc tactgtactt gaagaacgcg atccgcgtgg tctcggattc 4441 gcgcggcatc agcagccgga tcgtgcccga gcacgacgcc tccggcgccg cgctcgatct 4501 gtcgttcgcg cacctgcgca gcatctcgcc ctgccatctc gaatttctgc ggaacatggg 4561 cgtcagcgcc tcgatgtcgc tgtcgatcat cattgacggc acgctatggg gattgatcat 4621 ctgtcatcat tacgagccgc gtgccgtgcc gatggcgcag cgcgtcgcgg ccgaaatgtt 4681 cgccgacttc ttatcgctgc acttcaccgc cgcccaccac caacgctaag tcgacttgtt 4741 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc 4801 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttattcgac 4861 tgtttaaacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg 4921 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc 4981 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt 5041 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 5101 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt 5161 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc 5221 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa 5281 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa 5341 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc 5401 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc 5461 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag 5521 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga 5581 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc 5641 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac 5701 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg 5761 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca 5821 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa 5881 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa 5941 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt 6001 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag //