LOCUS       pEV715                  3874 bp    DNA     circular UNA 18-JAN-2021
DEFINITION  cHS4-2xgRNA1 BS-miniCMV1-EYFP-SV40.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
REFERENCE   1  (bases 1 to 3874)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
COMMENT     Imported From
            Path: C:\Users\UPALTO~1\AppData\Local\Temp\Rar$DRa0.846
            Filename: pL1F-1-s1-s2_RFP.ape
FEATURES             Location/Qualifiers
     source          1..3874
                     /mol_type="genomic DNA"
                     /organism="."
     misc_feature    1..45
                     /label=Seq2
                     /note="/ApEinfo_fwdcolor=#8000ff"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1..45
                     /label=seq2
                     /note="/ApEinfo_fwdcolor=#6666ff"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(40..45)
                     /label=BBb scar(4)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    54..71
                     /label=I-SceI
                     /note="/created by=User"
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    92..201
                     /label=RK2 trfa
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    206..211
                     /label=BBb scar
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     rep_origin      complement(229..911)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="/ApEinfo_fwdcolor=gray50"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_revcolor=gray50"
                     /note="color: #ffff00"
     misc_feature    827..832
                     /label=BBb scar(1)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(838..843)
                     /label=BBb scar(5)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    924..929
                     /label=BBb scar(2)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(936..941)
                     /label=BBb scar(6)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    1009..1866
                     /label=AmpR
                     /note="/ApEinfo_fwdcolor=#ff80ff"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1704..1709
                     /label=BBb scar(3)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(1721..1726)
                     /label=BBb scar(7)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    complement(1992..2009)
                     /label=I-SceI
                     /note="/created by=User"
                     /note="color: #31849b; direction: LEFT"
     misc_feature    2013..2057
                     /label=seq1
                     /note="/ApEinfo_fwdcolor=#8000ff"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    2058..2061
                     /label=GGAG
                     /note="/created by=elviravitu"
                     /note="/modified by=elviravitu"
                     /note="color: #31849b"
     misc_feature    2062..2591
                     /label=cHS4 Insulator x2
                     /note="Geneious type: chromatin insulator"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(2062..2591)
                     /label=Digestion of pZDonor-L_ISceI-Seq1-FseI-SeqX - F...
                     /note="Geneious type: Concatenated sequence"
                     /note="/NCBI Join Type=order"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    2062..2591
                     /label=Digestion of MAV1213-Hef1a-mKate2 - Fragment 1 ...
                     /note="Geneious type: Concatenated sequence"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2062..2591
                     /label=Digestion of MAV1212_Hef1a-mKate2 (reduced AarI...
                     /note="Geneious type: Concatenated sequence"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2323..2326
                     /label=AgeI
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2341..2591
                     /label=cHS4 Core
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    2612..2615
                     /label=TACT
                     /note="/created by=elviravitu"
                     /note="/modified by=elviravitu"
                     /note="color: #31849b"
     misc_feature    2616..2877
                     /label=pRJ_00030 (concatenated sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     rRNA            2665..2684
                     /label=gRNA1 BS
                     /note="/created by=User"
                     /note="/modified by=elviravitu"
                     /note="color: #00ff00; direction: RIGHT"
     misc_feature    2685..2687
                     /label=PAM
                     /note="Geneious type: motif"
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #a6acb3"
     rRNA            2690..2709
                     /label=gRNA1 BS
                     /note="/created by=User"
                     /note="/modified by=elviravitu"
                     /note="color: #00ff00; direction: RIGHT"
     misc_feature    2710..2712
                     /label=PAM
                     /note="Geneious type: motif"
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #a6acb3"
     misc_feature    2728..2744
                     /label=TALER 14 Bind
                     /note="Geneious type: Repressor"
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        2749..2808
                     /label=Minimal CMV
                     /note="/created by=User"
                     /note="color: #ffffff; direction: RIGHT"
     protein_bind    2749..2752
                     /label=TCTA
                     /note="/created by=elviravitu"
                     /note="color: #31849b"
     misc_feature    2817..2833
                     /label=TALER 14 Bind
                     /note="Geneious type: Repressor"
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2834..2850
                     /label=TALER 14 Bind
                     /note="Geneious type: Repressor"
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2851..2867
                     /label=TALER 14 Bind
                     /note="Geneious type: Repressor"
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    2878..2881
                     /label=AATG
                     /note="/created by=User"
                     /note="color: #31849b"
     unsure          2882..2920
                     /label=5' UTR
                     /note="/created by=elviravitu"
                     /note="color: #a6acb3"
     protein_bind    2921..2924
                     /label=AGGT
                     /note="/created by=User"
                     /note="color: #31849b"
     misc_feature    2925..2930
                     /label=Kozak
                     /note="Geneious type: motif"
                     /note="color: #a6acb3; direction: RIGHT"
     gene            2931..3650
                     /label=eYFP
                     /note="/modified by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(3430..3435)
                     /label=BBb scar(2)
                     /note="/ApEinfo_fwdcolor=cyan"
                     /note="/ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {}
                     0} width 5 offset 0"
                     /note="/ApEinfo_label=BBb scar"
                     /note="/ApEinfo_revcolor=green"
                     /note="color: #a6acb3; direction: LEFT"
     protein_bind    3651..3654
                     /label=GCTT
                     /note="/created by=User"
                     /note="color: #31849b"
     protein_bind    3675..3678
                     /label=CAAC
                     /note="/created by=User"
                     /note="color: #31849b"
     polyA_signal    3679..3870
                     /label=SV40 polyA
                     /note="/created by=User"
                     /note="/modified by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    3871..3874
                     /label=CGCT
                     /note="/created by=User"
                     /note="color: #31849b"
ORIGIN
        1 ggtgcgtttt tatgcttgta gtattgtata atgtttttaa gatccttaat taatagggat
       61 aacagggtaa tatcttgccc tgcacgaata ccagcgaccc cttgcccaaa tacttgccgt
      121 gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc tcctgcttgt
      181 cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg ccgcgttgct
      241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      661 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
      721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
      841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
      901 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
      961 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1021 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1081 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1141 accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1201 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1261 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1321 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1381 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1441 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1501 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1561 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1621 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1681 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1741 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1801 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     1861 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     1921 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacgaattg
     1981 gccagcgctg cattaccctg ttatccctaa ccggtttacc gagctcttat tggttttcaa
     2041 acttcattga ctgtgccgga ggcgcctggg agctcacggg gacagccccc ccccaaagcc
     2101 cccagggatg taattacgtc cctcccccgc tagggggcag cagcgagccg cccggggctc
     2161 cgctccggtc cggcgctccc cccgcatccc cgagccggca gcgtgcgggg acagcccggg
     2221 cacggggaag gtggcacggg atcgctttcc tctgaacgct tctcgctgct ctttgagcct
     2281 gcagacacct ggggggatac ggggaaaaag ctaggctgaa agaggttctt agaatgacac
     2341 gcgcctggga gctcacgggg acagcccccc cccaaagccc ccagggatgt aattacgtcc
     2401 ctcccccgct agggggcagc agcgagccgc ccggggctcc gctccggtcc ggcgctcccc
     2461 ccgcatcccc gagccggcag cgtgcgggga cagcccgggc acggggaagg tggcacggga
     2521 tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg cagacacctg gggggatacg
     2581 gggaaaaagc tggatcataa ctcgacgtgc atactgctcc gaatttctcg acagatctca
     2641 tgtgattacg ccaagctacg ggaggtcccc tccaccccac agtggggagg tcccctccac
     2701 cccacagtgg ggagcggaga ctctagatac ccctctccgc ttctgaattc taggcgtgta
     2761 cggtgggagg cctatataag cagagctcgt ttagtgaacc gtcagatcgc ctcgagtacc
     2821 cctctccgct tcttacccct ctccgcttct tacccctctc cgcttctgga tccaattaat
     2881 gctagtctac taccagagct catcgctagc gctaccggtc aggtgccacc atggtgagca
     2941 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     3001 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     3061 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     3121 ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     3181 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     3241 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     3301 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     3361 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     3421 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     3481 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     3541 accagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     3601 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga gcttccactg
     3661 gattgtacaa ttaccaacca gacatgataa gatacattga tgagtttgga caaaccacaa
     3721 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     3781 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     3841 aggttcaggg ggaggtgtgg gaggtttttt cgct
//
