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
//
