LOCUS       pJTM211_PCR_Product_-_pJM220-mNeongreen_not_codon_optimized_backbone_piece_-_P6d_PCR_Product_2_-_mNeonGreen        2986 bp    DNA     circular UNA 15-AUG-2022
DEFINITION  Gibson/Homology Cloning ligation product.
ACCESSION   urn.local.elinlarsson.10a-f5bxdjg
VERSION     urn.local.elinlarsson.10a-f5bxdjg
KEYWORDS    .
SOURCE      
  ORGANISM  .
FEATURES             Location/Qualifiers
     misc_feature    1..40
                     /standard_name="U1"
                     /label="U1"
     misc_feature    1..93
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:1wy-f5bxdlm"
                     /label="pJTM211 PCR Product extraction (concatenated
                     sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    1..40
                     /standard_name="U1 (concatenated sequence 1)"
                     /label="U1 (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     1..60
                     /standard_name="[1] U1-T7 (fwd) pJTM211"
                     /label="[1] U1-T7 (fwd) pJTM211"
     promoter        41..60
                     /standard_name="P_T7"
                     /label="P_T7"
     misc_feature    41..60
                     /standard_name="PT7 (concatenated sequence 2)"
                     /label="PT7 (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     RBS             61..93
                     /note="UTR1"
                     /standard_name="UTR1"
                     /label="UTR1"
     misc_feature    <61..>93
                     /standard_name
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    61..93
                     /standard_name="RBS (concatenated sequence 3)"
                     /label="RBS (concatenated sequence 3)"
                     /note="Geneious type: Concatenated sequence"
     ligation        71..93
                     /label="Ligation"
     misc_feature    71..841
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:105-f5bxdjg"
                     /label="pJM220-mNeongreen not codon optimized backbone
                     piece - P6d PCR Product 2 - mNeonGreen (concatenated
                     sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     complement(73..93)
                     /Sequence="TATATCTCCTTCTTAAAGTTA"
                     /Tm="45.8"
                     /%GC="23.8"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /created_by="elinlarsson"
                     /label="bb_R"
     CDS             94..822
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /label="mNeonGreen"
     primer          <94..107
                     /note="sequence:
                     actagtgctctgcagtaaggaggaacagctATGGTCTCCAAGGG"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="oGS32"
     ligation        94
                     /label="Ligation"
     misc_feature    <94..>822
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:29w-f5bxgmc"
                     /label="pJM220-mNeongreen not codon optimized backbone
                     piece (concatenated sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    94
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:29x-f5bxgmc"
                     /label="P6d PCR Product 2 (concatenated sequence 2)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     94..112
                     /annotation_group="334178000819817ec5a37aa-7c17-4c23-86cb-
                     df3d102e5c2e"
                     /Sequence="ATGGTCTCCAAGGGCGAGG"
                     /created_by="primer3"
                     /Product_Size=729
                     /#_Off-target_Sites=0
                     /Off-target_Template="pJM220-mNeongreen not codon
                     optimized backbone piece - P6d PCR Product 2 - mNeonGreen"
                     /Extension="TTTAACTTTAAGAAGGAGATATA"
                     /Tm_(binding_region)="61.4"
                     /Tm_(with_extension)="62.8"
                     /%GC_(binding_region)="63.2"
                     /%GC_(with_extension)="40.5"
                     /Hairpin_Tm_(with_extension)="50.3"
                     /Self_Dimer_Tm_(with_extension)="16.4"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="pJM220-mNeongreen not codon optimized backbone
                     piece - P6d PCR Product 2 - mNeonGreen (1 F) - pJTM211 PCR
                     Product"
     primer_bind     complement(776..835)
                     /created_by="elinlarsson"
                     /Sequence="TTTTTTGGCTGAATTAATGGTGATGGTGATGGTGCTTATAGAGCTCA
                     TCCATCCCCATCA"
                     /Tm="69.4"
                     /%GC="41.7"
                     /Hairpin_Tm="50.6"
                     /Self_Dimer_Tm="31.9"
                     /modified_by="elinlarsson"
                     /label="mNeongreen_R_overhang"
     primer_bind     complement(776..801)
                     /created_by="elinlarsson"
                     /Sequence="CTTATAGAGCTCATCCATCCCCATCA"
                     /Tm="61.5"
                     /%GC="46.2"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="17.8"
                     /modified_by="elinlarsson"
                     /label="mNeongreen_R"
     primer          complement(785..>822)
                     /note="sequence:
                     atcagaccgcttctgcgttctgattcgcgaTTACTTATAGAGCTCATCCA"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
                     /label="oGS33"
     primer_bind     complement(801..822)
                     /annotation_group="3341783828746837908cc99-9857-4125-9918-
                     987d5c4fab58"
                     /Sequence="TTAATGGTGATGGTGATGGTGC"
                     /created_by="primer3"
                     /Product_Size=729
                     /#_Off-target_Sites=0
                     /Off-target_Template="pJM220-mNeongreen not codon
                     optimized backbone piece - P6d PCR Product 2 - mNeonGreen"
                     /Extension="CTTAAGTTTTTTGGCTGAA"
                     /Tm_(binding_region)="59.0"
                     /Tm_(with_extension)="63.6"
                     /%GC_(binding_region)="45.5"
                     /%GC_(with_extension)="39.0"
                     /Hairpin_Tm_(with_extension)="42.1"
                     /Self_Dimer_Tm_(with_extension)="1.6"
                     /Pair_Dimer_Tm_(binding_region)="None"
                     /label="pJM220-mNeongreen not codon optimized backbone
                     piece - P6d PCR Product 2 - mNeonGreen (729 R) - pJTM211
                     PCR Product"
     misc_feature    802..819
                     /Original_Bases
                     /label
                     /note="Geneious type: Editing History Insertion"
     misc_feature    802..819
                     /label="his tag"
     terminator      823..912
                     /standard_name="ECK120029600 (strongest natural Voigt,
                     378x)"
                     /label="ECK120029600 (strongest natural Voigt, 378x)"
     ligation        823..841
                     /label="Ligation"
     misc_feature    823..2986
                     /Source="referenceToDocument:
                     urn:local:elinlarsson:1wy-f5bxdlm"
                     /label="pJTM211 PCR Product extraction (concatenated
                     sequence 3)"
                     /note="Geneious type: Concatenated sequence"
     misc_feature    823..912
                     /standard_name="T12m (concatenated sequence 5)"
                     /label="T12m (concatenated sequence 5)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     823..846
                     /created_by="elinlarsson"
                     /Sequence="TTCAGCCAAAAAACTTAAGACCGC"
                     /Tm="60.7"
                     /%GC="41.7"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="10.1"
                     /modified_by="elinlarsson"
                     /label="bb_F"
     primer_bind     complement(885..944)
                     /standard_name="[6] T12m-UX (rev)"
                     /label="[6] T12m-UX (rev)"
     misc_feature    913..952
                     /standard_name="UX"
                     /label="UX"
     misc_feature    913..952
                     /standard_name="UX (concatenated sequence 6)"
                     /label="UX (concatenated sequence 6)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     919..978
                     /standard_name="[7] UX-pUC19 (fwd)"
                     /label="[7] UX-pUC19 (fwd)"
     source          join(2634..>2986,<953..2633)
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     misc_feature    953..2986
                     /standard_name="pUC19 Linear (concatenated sequence 7)"
                     /label="pUC19 Linear (concatenated sequence 7)"
                     /note="Geneious type: Concatenated sequence"
     primer_bind     953..972
                     /standard_name="pUC19-linear-fwd"
                     /label="pUC19-linear-fwd"
     promoter        1126..1230
                     /gene="bla"
                     /standard_name="AmpR promoter"
                     /label="AmpR promoter"
     CDS             1231..2091
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /standard_name="AmpR"
                     /label="AmpR"
     rep_origin      2262..2850
                     /label="rep origin"
     primer_bind     complement(2969..2986)
                     /standard_name="pUC19-linear-rev"
                     /label="pUC19-linear-rev"
ORIGIN      
        1 cattactcgc atccattctc aggctgtctc gtctcgtctc taatacgact cactataggg
       61 aataattttg tttaacttta agaaggagat ataatggtct ccaagggcga ggaagataat
      121 atggcgtccc tccccgcgac gcatgaactc catatcttcg ggtcgatcaa cggggtggac
      181 tttgatatgg tggggcaagg gacgggcaac cccaatgatg gctacgaaga gctcaacctc
      241 aagagcacga agggcgacct gcagtttagc ccctggatcc tggtgccgca catcgggtat
      301 ggcttccatc aatatctgcc ctaccccgac gggatgtccc cgtttcaggc ggccatggtc
      361 gacggctcgg ggtaccaagt gcatcgcacg atgcagtttg aggatggcgc ctccctgacc
      421 gtgaactatc gctacacgta cgaaggcagc catatcaagg gggaagcgca ggtgaagggc
      481 accggctttc cggcggacgg gccggtgatg accaattccc tgacggccgc ggattggtgt
      541 cggtccaaga agacgtaccc gaacgataag accatcatca gcacgttcaa gtggtcgtat
      601 accaccggca acggcaagcg ctaccgttcg acggcccgca cgacgtacac ctttgccaag
      661 ccgatggcgg cgaactatct gaagaaccaa cccatgtatg tgttccggaa gaccgagctc
      721 aagcattcga agacggagct gaattttaag gaatggcaga aggcgtttac cgatgtgatg
      781 gggatggatg agctctataa gcaccatcac catcaccatt aattcagcca aaaaacttaa
      841 gaccgccggt cttgtccact accttgcagt aatgcggtgg acaggatcgg cggttttctt
      901 ttctcttctc aaccaggata catagattac cacaactccg agcccttcca ccccgcttac
      961 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     1021 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1081 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1141 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1201 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     1261 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     1321 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     1381 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     1441 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     1501 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1561 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1621 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1681 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1741 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1801 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1861 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1921 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     1981 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     2041 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     2101 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2161 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2221 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2281 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2341 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2401 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2461 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2521 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2581 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2641 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2701 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2761 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2821 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2881 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2941 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgca
//
