LOCUS       Exported                3261 bp ds-DNA     circular SYN 14-OCT-2022
DEFINITION  A new nucleotide sequence entered manually.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3261)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 18, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3261
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..105
                     /gene="<i>bla</i>"
                     /label=AmpR promoter
                     /note="Derived using Geneious Prime 2021.2.1 'Annotate from
                     Database' based on nucleotide similarity"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a 
                     href=""urn:local:.:32o-d8wha7z"">pCB170_hU6-k87(DR30)_EF1a-
                     k87Cas13d-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a href=""urn:local:.:x-czcif9y"">Pseudosite
                     attD 56/10 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a 
                     href=""urn:local:.:10-d4l9cgg"">Pseudosite attD 56/17 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a 
                     href=""urn:local:.:2ll-d8wh8yi"">pCB169_Inducible 
                     k87-msfGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a 
                     href=""urn:local:.:341-d8wha9z"">pCB189_PB_inducible_Adm-NL
                     S-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a 
                     href=""urn:local:.:34a-d8whaae"">pCB192_PB_inducible_twins_
                     985-NES-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a 
                     href=""urn:local:.:zb-c9vfl3m"">pC439_EF1a-PhiC31(optimized
                     )-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a 
                     href=""urn:local:.:2c-de59zgs"">pCB214_pRec_(no 
                     attP)_mcherry-pA</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR promoter (<a href=""urn:local:.:m-czcg5p2"">Pseudosite
                     attD new20/3</a>)"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; 
                     AmpR promoter (<a 
                     href=""urn:local:.:1y-czcnqw5"">Pseudosite attD 56/30 
                     Puro</a>)"
                     /note="/Transferred From=Pseudosite attD 56/30 Puro"
                     /note="/Transferred Similarity=100.00%"
     CDS             106..966
                     /standard_name="AmpR"
                     /label=AmpR
                     /note="Derived using Geneious Prime 2021.2.1 'Annotate from
                     Database' based on nucleotide similarity"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:10w-c9vfrsv"">pC457</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:1a2-dcuz67h"">pC468</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:10z-c9vfrsy"">pC458</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:1bb-dcuz682"">pC483</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:15-cnpec3p"">pC497</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:118-c9vfrt7"">pC461</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:12b-c9vfru9"">pC474</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:10t-c9vfrsr"">pC456</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     AmpR (<a href=""urn:local:.:12e-c9vfruc"">pC475</a>)"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; 
                     AmpR (<a href=""urn:local:.:115-c9vfrt4"">pC460</a>)"
                     /note="/Transferred From=pC460"
                     /note="/Transferred Similarity=100.00%"
     rep_origin      1137..1725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="Derived using Geneious Prime 2021.2.1 'Annotate from
                     Database' based on nucleotide similarity"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a 
                     href=""urn:local:.:32o-d8wha7z"">pCB170_hU6-k87(DR30)_EF1a-
                     k87Cas13d-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a href=""urn:local:.:x-czcif9y"">Pseudosite attD 
                     56/10 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a href=""urn:local:.:10-d4l9cgg"">Pseudosite attD 
                     56/17 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a 
                     href=""urn:local:.:zb-c9vfl3m"">pC439_EF1a-PhiC31(optimized
                     )-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a href=""urn:local:.:2c-de59zgs"">pCB214_pRec_(no 
                     attP)_mcherry-pA</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a href=""urn:local:.:m-czcg5p2"">Pseudosite attD 
                     new20/3</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a href=""urn:local:.:1a-d4ladew"">Pseudosite attD 
                     Phic31 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a href=""urn:local:.:22-czco819"">Pseudosite attD 
                     Bxb1 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     ori (<a href=""urn:local:.:2e-czcp600"">Pseudosite attD 
                     Pa01 Puro</a>)"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; ori
                     (<a href=""urn:local:.:1y-czcnqw5"">Pseudosite attD 56/30 
                     Puro</a>)"
                     /note="/Transferred From=Pseudosite attD 56/30 Puro"
                     /note="/Transferred Similarity=100.00%"
                     /note="/snapgene_direction=RIGHT"
     promoter        1910..3087
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
                     /note="Derived using Geneious Prime 2021.2.1 'Annotate from
                     Database' based on nucleotide similarity"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:32o-d8wha7z"">pCB170_hU6-k87(DR30)_EF1a-
                     k87Cas13d-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:30g-d8wha45"">pCB144_PB_Constitutive_Cas
                     Rx-msfGFP_v2_(Use_me!)</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:x-czcif9y"">Pseudosite attD 56/10 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:10-d4l9cgg"">Pseudosite attD 56/17 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:zb-c9vfl3m"">pC439_EF1a-PhiC31(optimized
                     )-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:m-czcg5p2"">Pseudosite attD 
                     new20/3</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:1a-d4ladew"">Pseudosite attD Phic31 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:22-czco819"">Pseudosite attD Bxb1 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:2jj-d8wh8ul"">pCB144_PB_Constitutive_Cas
                     Rx-msfGFP_v2_(Use_me!)</a>)"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; 
                     EF-1-alpha promoter (<a 
                     href=""urn:local:.:1y-czcnqw5"">Pseudosite attD 56/30 
                     Puro</a>)"
                     /note="/Transferred From=Pseudosite attD 56/30 Puro"
                     /note="/Transferred Similarity=100.00%"
     intron          2141..3078
                     /label=EF-1-alpha intron A
                     /note="Derived using Geneious Prime 2021.2.1 'Annotate from
                     Database' based on nucleotide similarity"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     EF-1-alpha intron A (<a 
                     href=""urn:local:.:ha-dpuu0sj"">pJT301_EF1a-attB-BxB1(</a>)
                     "
                     /note="/ApEinfo_fwdcolor=#85dae9"
                     /note="/ApEinfo_revcolor=#85dae9"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; 
                     EF-1-alpha intron A (<a 
                     href=""urn:local:.:gr-dputnng"">pJT301_EF1a-attB-BxB1 
                     Landing pad final</a>)"
                     /note="/Transferred From=pJT301_EF1a-attB-BxB1 Landing pad 
                     final"
                     /note="/Transferred Similarity=100.00%"
     misc_feature    3088..3199
                     /label=Si74 attB
                     /note="/created by=aliso"
     regulatory      3203..3212
                     /regulatory_class="other"
                     /standard_name="Kozak sequence"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for
                     strong initiation of translation (<a 
                     href=""https://www.ncbi.nlm.nih.gov/pubmed/3313277"" 
                     title=""https://www.ncbi.nlm.nih.gov/pubmed/3313277"">Koza 
                     k, 1987</a>)"
                     /note="Derived using Geneious Prime 2021.2.1 'Annotate from
                     Database' based on nucleotide similarity"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:2cb-d8wh8nm"">pCB046_pAAV-U6-DR30-BsaI-E
                     FS-hnRNPA1-dCasRx-bghpA</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:30g-d8wha45"">pCB144_PB_Constitutive_Cas
                     Rx-msfGFP_v2_(Use_me!)</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:2ll-d8wh8yi"">pCB169_Inducible 
                     k87-msfGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:341-d8wha9z"">pCB189_PB_inducible_Adm-NL
                     S-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:34a-d8whaae"">pCB192_PB_inducible_twins_
                     985-NES-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:2c-de59zgs"">pCB214_pRec_(no 
                     attP)_mcherry-pA</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:2on-d8wh92t"">pCB208_PB_inducible_dCas9-
                     KRAB-MeCP2-2A-EGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:32i-d8wha7s"">pCB169_Inducible 
                     k87-msfGFP</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     Kozak sequence (<a 
                     href=""urn:local:.:2jj-d8wh8ul"">pCB144_PB_Constitutive_Cas
                     Rx-msfGFP_v2_(Use_me!)</a>)"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; 
                     Kozak sequence (<a 
                     href=""urn:local:.:30j-d8wha48"">pCB145_PB_Inducible_CasRx-
                     msfGFP_v3_(Use_me!)</a>)"
                     /note="/Transferred 
                     From=pCB145_PB_Inducible_CasRx-msfGFP_v3_(Use_me!)"
                     /note="/Transferred Similarity=100.00%"
     CDS             3209..3231
                     /codon_start=1
                     /transl_table=0
                     /product="<html><body>enhanced monomeric blue fluorescent 
                     protein (<a 
                     href=""http://www.ncbi.nlm.nih.gov/pubmed/22174863"" 
                     title=""http://www.ncbi.nlm.nih.gov/pubmed/22174863"">Subac
                     h et al., 2011</a>)</body></html>"
                     /label=Stop codons
                     /note="<html><body>mammalian codon-optimized</body></html>"
                     /note="Derived using Geneious Prime 2021.2.1 'Annotate from
                     Database' based on nucleotide similarity"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mTagBFP2 (<a href=""urn:local:.:v-d6qnnp5"">attA 
                     pCB237_pRec_EF1a-(no attB)_STOP_pA</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mTagBFP2 (<a 
                     href=""urn:local:.:2v-daaegtu"">pCB237_pRec_EF1a-(no 
                     attB)_STOP_pA</a>)"
                     /note="/Original 
                     Translation=MVSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRI
                     KVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVL
                     TATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMA
                     LKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVAR
                     YCDLPSKLGHKLN"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; 
                     mTagBFP2 (<a href=""urn:local:.:w-cquvphe"">Truncated Pa01 
                     attB (34)</a>)"
                     /note="/Transferred From=Truncated Pa01 attB (34)"
                     /note="/Transferred Similarity=100.00%"
                     /note="/modified by=aliso"
                     /translation="MG***G"
ORIGIN
        1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
       61 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg gctccggtgc
     1921 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     1981 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2041 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     2101 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
     2161 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccactgg
     2221 ctgcagtacg tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg
     2281 ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg
     2341 ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct
     2401 ctagccattt aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt
     2461 gtaaatgcgg gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac
     2521 ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg
     2581 agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg
     2641 ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg
     2701 gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg
     2761 ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc
     2821 gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc
     2881 ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac
     2941 actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa
     3001 tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt
     3061 ttttttcttc catttcaggt gtcgtgattc cgacgcagtt tccgacgagt acgaggacga
     3121 ggacagacgt gcctaccggc aaggtcaagt ggttcaacag cgagaagggc ttcggctttc
     3181 tctcccgcga cgacggcggg ccgccaccat gggatagtga taaggatagt gacgtcaggt
     3241 ggcacttttc ggggaaatgt g
//
