LOCUS       Exported                3025 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 3025)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Oct 18, 2022 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3025
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        32..136
                     /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             137..997
                     /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      1168..1756
                     /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"
     misc_feature    1951..2062
                     /label=Si74 attP
                     /note="/created by=aliso"
     regulatory      2070..2079
                     /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             2076..2786
                     /codon_start=1
                     /transl_table=0
                     /gene="mCherry"
                     /product="red fluorescent protein mCherry"
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein"
                     /note="codon-optimized for expression in mammalian cells"
                     /note="/dnas_title=mCherry"
                     /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;
                     mCherry (<a href=""urn:local:.:15-cnpec3p"">pC497</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:x-czcif9y"">Pseudosite attD 
                     56/10 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:10-d4l9cgg"">Pseudosite attD
                     56/17 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:2c-de59zgs"">pCB214_pRec_(no
                     attP)_mcherry-pA</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:m-czcg5p2"">Pseudosite attD 
                     new20/3</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:1a-d4ladew"">Pseudosite attD
                     Phic31 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:22-czco819"">Pseudosite attD
                     Bxb1 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:2e-czcp600"">Pseudosite attD
                     Pa01 Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     mCherry (<a href=""urn:local:.:bx-cm0clbm"">Pseudosite attD
                     new 20/20</a>)"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; 
                     mCherry (<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="/Transferred 
                     Translation=MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFED
                     GGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGE
                     IKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAEG
                     RHSTGGMDELYK*"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     polyA_signal    2817..3024
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /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;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:x-czcif9y"">Pseudosite attD 56/10 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:10-d4l9cgg"">Pseudosite attD 56/17 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:2c-de59zgs"">pCB214_pRec_(no 
                     attP)_mcherry-pA</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:m-czcg5p2"">Pseudosite attD 
                     new20/3</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:1a-d4ladew"">Pseudosite attD Phic31 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:2te-d8wh9xl"">pAAV-EF1a-hChR2-EYFP-WPRE<
                     /a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:22-czco819"">Pseudosite attD Bxb1 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:2e-czcp600"">Pseudosite attD Pa01 
                     Puro</a>)"
                     /note="/Alternative Match=100.00%; Adjusted-Interval-0:0/0;
                     bGH poly(A) signal (<a 
                     href=""urn:local:.:bx-cm0clbm"">Pseudosite attD new 
                     20/20</a>)"
                     /note="/Primary Match=100.00%; Adjusted-Interval-0:0/0; bGH
                     poly(A) signal (<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%"
ORIGIN
        1 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
       61 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
      121 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
      181 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
      241 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
      301 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
      361 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
      421 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
      481 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
      541 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
      601 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
      661 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
      721 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
      781 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
      841 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
      901 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
      961 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     1021 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     1081 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     1141 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     1201 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     1261 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     1321 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     1381 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     1441 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     1501 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     1561 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     1621 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     1681 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     1741 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     1801 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     1861 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     1921 gagcgaggaa gcggaagagc agagacgcgt tagtgacgtc tgtccgcgca gtgatcgagg
     1981 gagtgtgtgc tttgccgact ggcaaggtca agccggtctg ctaggcacag agagccggta
     2041 cagtcctccc catgcaaccc aactccgccg ccaccatggt gagcaagggc gaggaggata
     2101 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     2161 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2221 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     2281 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     2341 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     2401 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     2461 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2521 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     2581 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     2641 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     2701 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2761 gcggcatgga cgagctgtac aagtaactcg agctagagct cgctgatcag cctcgactgt
     2821 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2881 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2941 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3001 agacaatagc aggcatgctg gggac
//
