LOCUS       pEV785                  5192 bp    DNA     circular SYN 14-DEC-2020
DEFINITION  cHS4-2xgRNA1_BS-YB_TATA-1xuORF-Kozak3-Gal4-VPR-polyA(SV40).
ACCESSION   urn.local...1sj-cjz3hzz
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5192)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
COMMENT     Imported From
            Path: /Users/elviravitu/Dropbox (MIT)/Elvira 
            collaboration/Plasmids/Resheaper library
            Filename: 10 documents from Reshpaer first cloning round.gb
FEATURES             Location/Qualifiers
     source          1..5192
                     /lab_host="."
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1802..1834
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     misc_feature    13..18
                     /label=BBb scar
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(25..30)
                     /label=BBb scar
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    98..955
                     /label=AmpR
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    793..798
                     /label=BBb scar
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(810..815)
                     /label=BBb scar
                     /note="color: #a6acb3; direction: LEFT"
     promoter        complement(997..1025)
                     /label=amp prom
                     /note="color: #ffffff; direction: LEFT"
     misc_feature    1102..1146
                     /label=seq1
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1147..1735
                     /label=pBjh_3260 (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1147..1161
                     /label=pL0 I
                     /note="Geneious type: gg_overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1147..1161
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1147..1161
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1147..1161
                     /label=ligation
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1162..1731
                     /label=2x cHS4 Insulator
                     /note="Geneious type: Insulator"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    1240..1244
                     /label=CTCF Bind (Short)
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    1519..1523
                     /label=CTCF Bind (Short)
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    1732..1838
                     /label=pEV700 (concatenated sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     rRNA            1736..1755
                     /label=gRNA1 BS
                     /note="/created_by=User"
                     /note="/modified_by=elviravitu"
                     /note="color: #00ff00; direction: RIGHT"
     misc_feature    1756..1758
                     /label=PAM
                     /note="Geneious type: motif"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     rRNA            1777..1796
                     /label=gRNA1 BS
                     /note="/created_by=User"
                     /note="/modified_by=elviravitu"
                     /note="color: #00ff00; direction: RIGHT"
     misc_feature    1797..1799
                     /label=PAM
                     /note="Geneious type: motif"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1800..1834
                     /label=Promoter
                     /note="Geneious type: motif"
                     /note="/created_by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        1810..1834
                     /label=YB_TATA
                     /note="/created_by=elviravitu"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    1835..1898
                     /label=pEV711 (concatenated sequence 4)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     unsure          1839..1877
                     /label=5' UTR
                     /note="/created by=elviravitu"
                     /note="color: #a6acb3"
     misc_feature    1878..1894
                     /label=Digestion of 13.58 - Fragment 2 extraction (con...
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1884..1895
                     /label=uORF x1
                     /note="/created by=User"
                     /note="/modified by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1899..4050
                     /label=pEV740 (concatenated sequence 5)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1899..2354
                     /label=pEV720 (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1899..1908
                     /label=Kozak3
                     /note="Geneious type: motif"
                     /note="/created_by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             1905..1928
                     /label=Gal4-NLS-VPR
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #993366; direction: RIGHT"
     CDS             1929..2354
                     /label=Gal4
                     /note="Geneious type: gene"
                     /note="/created_by=User"
                     /note="/modified by=elviravitu"
                     /note="/modified_by=User"
                     /note="/modified_by=elviravitu"
                     /note="color: #993366; direction: RIGHT"
     misc_feature    2351..4050
                     /label=pEV731 (concatenated sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2351..2354
                     /label=GGGC
                     /note="Geneious type: Q"
                     /note="/created_by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2351..2354
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2355..2375
                     /label=SV40 NLS
                     /note="Geneious type: motif"
                     /note="/Transferred_From=Digestion of pRJ_00009 PCR Product
                     - Fragment 2, Digestion of pRJ_00010 PCR Product - Fragment
                     2, Digestion of pRJ_00011 PCR Product - Fragment 2, 
                     Digestion of pRJ_00012 PCR Product - Fragment 2, Digestion 
                     of pRJ_00013 PCR Product - Fragment 2, pRJ_00009, pRJ_00009
                     PCR Product, pRJ_00010, pRJ_00010 PCR Product, pRJ_00011, 
                     pRJ_00011 PCR Product, pRJ_00012, pRJ_00012 PCR Product, 
                     pRJ_00013, pRJ_00013 PCR Product, pRJ_00095, pRJ_00096, 
                     pRJ_00097, pRJ_00098, pRJ_00099"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2376..2384
                     /label=GSG
                     /note="Geneious type: motif"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2422..2442
                     /label=attB1
                     /note="Geneious type: recomb_site"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2469..2636
                     /label=VP64
                     /note="Geneious type: domain"
                     /note="/Transferred_From=Digestion of pRJ_00010 PCR Product
                     - Fragment 2, Digestion of pRJ_00011 PCR Product - Fragment
                     2, pRJ_00010, pRJ_00010 PCR Product, pRJ_00011, pRJ_00011 
                     PCR Product, pRJ_00096, pRJ_00097"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2649..2672
                     /label=SV40
                     /note="Geneious type: motif"
                     /note="/Transferred_From=Digestion of pRJ_00010 PCR Product
                     - Fragment 2, pRJ_00010, pRJ_00010 PCR Product, pRJ_00096"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2673..3455
                     /label=p65
                     /note="Geneious type: domain"
                     /note="/Transferred_From=Digestion of pRJ_00010 PCR Product
                     - Fragment 2, Digestion of pRJ_00012 PCR Product - Fragment
                     2, pRJ_00010, pRJ_00010 PCR Product, pRJ_00012, pRJ_00012 
                     PCR Product, pRJ_00096, pRJ_00098"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2718..2729
                     /label=NLS
                     /note="Geneious type: motif"
                     /note="/Transferred_From=p65 (pBjh_4067)"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3060..3194
                     /label=Activation domain
                     /note="Geneious type: Region_of_interest"
                     /note="/Transferred_From=p65 (pBjh_4067)"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3456..3473
                     /label=GS-rich Linker
                     /note="Geneious type: motif"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3474..4043
                     /label=Rta
                     /note="Geneious type: domain"
                     /note="/Transferred_From=Digestion of pRJ_00010 PCR Product
                     - Fragment 2, Digestion of pRJ_00013 PCR Product - Fragment
                     2, pRJ_00010, pRJ_00010 PCR Product, pRJ_00013, pRJ_00013 
                     PCR Product, pRJ_00096, pRJ_00099"
                     /note="/Transferred_Similarity=100.00%"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    4047..4074
                     /label=pEV542 (concatenated sequence 6)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     unsure          4047..4050
                     /label=Ligation
                     /note="/created_by=fabio"
                     /note="color: #a6acb3"
     misc_feature    4047..4050
                     /label=L0.G_Backbone (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/NCBI Join Type=join"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    4071..4270
                     /label=pEV618 (concatenated sequence 7)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     polyA_signal    4075..4266
                     /label=SV40 polyA
                     /note="/created by=User"
                     /note="/modified by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    4267..6342
                     /label=L1.Seq1-Seq2_Backbone (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    4282..4326
                     /label=seq2
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(4321..4326)
                     /label=BBb scar
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    4373..4482
                     /label=RK2 trfa
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    4487..4492
                     /label=BBb scar
                     /note="color: #a6acb3; direction: RIGHT"
     rep_origin      complement(4510..5192)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="color: #ffff00"
     misc_feature    5108..5113
                     /label=BBb scar
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    complement(5119..5124)
                     /label=BBb scar
                     /note="color: #a6acb3; direction: LEFT"
ORIGIN
        1 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
       61 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      121 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      181 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagaac
      241 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      301 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      361 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      421 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      481 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      541 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      601 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
      661 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
      721 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
      781 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
      841 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
      901 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
      961 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1021 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacgaattgg ccagcgctgc
     1081 attaccctgt tatccctaac cggtttaccg agctcttatt ggttttcaaa cttcattgac
     1141 tgtgccgctc ttcaaccgga gttctgcaat tcatgtgatc tgcgcctggg agctcacggg
     1201 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     1261 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     1321 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     1381 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ctaggctgaa
     1441 agaggatctt agaatgacac gcgcctggga gctcacgggg acagcccccc cccaaagccc
     1501 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     1561 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     1621 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     1681 cagacacctg gggggatacg gggaaaaagc tggatcataa ctcgacgtgc atactgtccc
     1741 ctccacccca cagtggggca tagcgccggc cgcgccgtcc cctccacccc acagtggggc
     1801 accatatgct ctagagggta tataatgggg gccaaatgct agtctactac cagagctcat
     1861 cgctagcgct accggtcttt gtaaccatgg gttgaggtgc ctttatggtg aagctactgt
     1921 cctctatcga acaagcatgc gatatttgcc gacttaaaaa gctcaagtgc tccaaagaaa
     1981 aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg tcgctactct cccaaaacca
     2041 aaaggtcccc gctgactagg gcacatctga cagaagtgga atcaaggcta gaaagactgg
     2101 aacagctatt tctactgatt tttcctcgag aggaccttga catgattttg aaaatggatt
     2161 ctttacagga tataaaagca ttgttaacag gattatttgt acaagataat gtgaataaag
     2221 atgccgtcac agatagattg gcttcagtgg agactgatat gcctctaaca ttgagacagc
     2281 atagaataag tgcgacatca tcatcggaag agagtagtaa caaaggtcaa agacagttga
     2341 ctgtatcgat gggccccaag aagaagagga aggtgggatc tggctcgcca gggatccgtc
     2401 gacttgacgc gttgatatca acaagtttgt acaaaaaagc aggctacaaa gaggccagcg
     2461 gttccggacg ggctgacgca ttggacgatt ttgatctgga tatgctggga agtgacgccc
     2521 tcgatgattt tgaccttgac atgcttggtt cggatgccct tgatgacttt gacctcgaca
     2581 tgctcggcag tgacgccctt gatgatttcg acctggacat gctgattaac tctagaagtt
     2641 ccggatctcc gaaaaagaaa cgcaaagttg gtagccagta cctgcccgac accgacgacc
     2701 ggcaccggat cgaggaaaag cggaagcgga cctacgagac attcaagagc atcatgaaga
     2761 agtccccctt cagcggcccc accgacccta gacctccacc tagaagaatc gccgtgccca
     2821 gcagatccag cgccagcgtg ccaaaacctg ccccccagcc ttaccccttc accagcagcc
     2881 tgagcaccat caactacgac gagttcccta ccatggtgtt ccccagcggc cagatctctc
     2941 aggcctctgc tctggctcca gcccctcctc aggtgctgcc tcaggctcct gctcctgcac
     3001 cagctccagc catggtgtct gcactggctc aggcaccagc acccgtgcct gtgctggctc
     3061 ctggacctcc acaggctgtg gctccaccag cccctaaacc tacacaggcc ggcgagggca
     3121 cactgtctga agctctgctg cagctgcagt tcgacgacga ggatctggga gccctgctgg
     3181 gaaacagcac cgatcctgcc gtgttcaccg acctggccag cgtggacaac agcgagttcc
     3241 agcagctgct gaaccagggc atccctgtgg cccctcacac caccgagccc atgctgatgg
     3301 aataccccga ggccatcacc cggctcgtga caggcgctca gaggcctcct gatccagctc
     3361 ctgcccctct gggagcacca ggcctgccta atggactgct gtctggcgac gaggacttca
     3421 gctctatcgc cgatatggat ttctcagcct tgctgggctc tggcagcggc agccgggatt
     3481 ccagggaagg gatgtttttg ccgaagcctg aggccggctc cgctattagt gacgtgtttg
     3541 agggccgcga ggtgtgccag ccaaaacgaa tccggccatt tcatcctcca ggaagtccat
     3601 gggccaaccg cccactcccc gccagcctcg caccaacacc aaccggtcca gtacatgagc
     3661 cagtcgggtc actgaccccg gcaccagtcc ctcagccact ggatccagcg cccgcagtga
     3721 ctcccgaggc cagtcacctg ttggaggatc ccgatgaaga gacgagccag gctgtcaaag
     3781 cccttcggga gatggccgat actgtgattc cccagaagga agaggctgca atctgtggcc
     3841 aaatggacct ttcccatccg cccccaaggg gccatctgga tgagctgaca accacacttg
     3901 agtccatgac cgaggatctg aacctggact cacccctgac cccggaattg aacgagattc
     3961 tggatacctt cctgaacgac gagtgcctct tgcatgccat gcatatcagc acaggactgt
     4021 ccatcttcga cacatctctg ttttgagctt ccactggatt gtacaattac caaccagaca
     4081 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4141 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4201 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     4261 ttttttcgct cgctgaagag cggtgcgttt ttatgcttgt agtattgtat aatgttttta
     4321 agatccttaa ttaataggga taacagggta atatcttgcc ctgcacgaat accagcgacc
     4381 ccttgcccaa atacttgccg tgggcctcgg cctgagagcc aaaacacttg atgcggaaga
     4441 agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca catctaggat ctgccaggaa
     4501 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4561 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4621 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4681 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4741 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4801 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4861 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4921 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4981 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5041 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5101 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5161 cgaaaactca cgttaaggga ttttggtcat ga
//
