LOCUS       pEV749                  3809 bp    DNA     circular SYN 11-DEC-2020
DEFINITION  cHS4-5xUAS (spacer)-YB_TATA-EYFP.
ACCESSION   urn.local...1a-ck48kjp
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3809)
  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..3809
                     /lab_host="."
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          2784..2816
                     /mol_type="other DNA"
                     /note="color: #ffffff"
                     /organism="synthetic DNA construct"
     misc_feature    1..2087
                     /label=ST2-3 (concatenated sequence 1)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     primer_bind     1..43
                     /label=RJ_00429Fc
                     /note="color: black; sequence: 
                     CGGTAATGGTCTCACGCTTACTGAAGAGCGGTCTAATACCCAATCTCTCGTCTTATC"
     misc_feature    1..4
                     /label=pL0 T
                     /note="Geneious type: Gg_overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1..4
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1
                     /label=Infusion
                     /note="Geneious type: Gibson_Overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     misc_feature    5..7
                     /label=pL1 T3
                     /note="Geneious type: Gg_overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    complement(9..15)
                     /label=SapI Bind
                     /note="/created_by=User"
                     /note="color: #31849b; direction: LEFT"
     misc_feature    16..60
                     /label=pL1 S3
                     /note="Geneious type: Gibson_Overhang"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     rep_origin      107..216
                     /direction=RIGHT
                     /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: #ffff00"
     primer_bind     complement(213..232)
                     /label=JH1030R
                     /note="color: black; sequence: CCTGGCAGATCCTAGATGTG"
     rep_origin      complement(244..926)
                     /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"
     CDS             complement(1021..1881)
                     /label=AmpR
                     /note="/created_by=User"
                     /note="color: #993366; direction: LEFT"
     primer_bind     1712..1732
                     /label=JH1030F
                     /note="color: black; sequence: CTCTCAAGGATCTTACCGCTG"
     misc_feature    complement(1882..1895)
                     /label=RBS
                     /note="Geneious type: motif"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: LEFT"
     primer_bind     1916..1936
                     /label=JH856R
                     /note="color: black; sequence: CAGGGTTATTGTCTCATGAGC"
     misc_feature    complement(1923..1951)
                     /label=pAmp
                     /note="Geneious type: promoter_prokaryotic"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: LEFT"
     misc_feature    2028..2072
                     /label=pL1 S2
                     /note="Geneious type: Gibson_Overhang"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     primer_bind     complement(2032..2088)
                     /label=RJ_00429Rc
                     /note="color: black; sequence: 
                     GTGATACGGTCTCACTCCGCGTGAAGAGCGGATCTTAAAAACATTATACAATACTACAA
                     GCATAAAAACG"
     protein_bind    2073..2079
                     /label=SapI Bind
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    2081..2083
                     /label=pL1 T2
                     /note="Geneious type: Gg_overhang"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2084..2661
                     /label=pBjh_3260 (concatenated sequence 2)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2084..2087
                     /label=pL0 I
                     /note="Geneious type: Gg_overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2084..2087
                     /label=pL0 I
                     /note="Geneious type: gg_overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2084..2087
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2084..2087
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2087
                     /label=Infusion
                     /note="Geneious type: Gibson_Overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3"
     misc_feature    2088..2657
                     /label=2x cHS4 Insulator
                     /note="Geneious type: Insulator"
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    2166..2170
                     /label=CTCF Bind (Short)
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
     protein_bind    2445..2449
                     /label=CTCF Bind (Short)
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
     misc_feature    2658..2820
                     /label=pEV729 (concatenated sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2658..2661
                     /label=TACT
                     /note="Geneious type: polylinker"
                     /note="/created_by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2658..2661
                     /label=pL0 I-P
                     /note="Geneious type: gg_overhang"
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2658..2661
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2658..2661
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2662..2778
                     /label=pRJ_00030 (concatenated sequence 3)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     enhancer        2662..2678
                     /label=UAS (Gal4)
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     enhancer        2687..2703
                     /label=UAS (Gal4)
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     enhancer        2712..2728
                     /label=UAS (Gal4)
                     /note="/created_by=User"
                     /note="/modified_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     enhancer        2737..2753
                     /label=UAS (Gal4)
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     enhancer        2762..2778
                     /label=UAS (Gal4)
                     /note="/created_by=User"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2782..2816
                     /label=Promoter
                     /note="Geneious type: motif"
                     /note="/created_by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     promoter        2792..2816
                     /label=YB_TATA
                     /note="/created_by=elviravitu"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    2817..2863
                     /label=pEV711 (concatenated sequence 4)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    2817..2820
                     /label=AATG
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
     unsure          2821..2859
                     /label=5' UTR
                     /note="/created_by=elviravitu"
                     /note="color: #a6acb3"
     misc_feature    2860..3593
                     /label=L0.G_EYFP (concatenated sequence 5)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    2864..2869
                     /label=Kozak
                     /note="Geneious type: motif"
                     /note="color: #a6acb3; direction: RIGHT"
     ncRNA           2870..3589
                     /label=eYFP
                     /note="/modified_by=User"
                     /note="color: #8fbc8f; direction: RIGHT"
     primer_bind     3169..3183
                     /label=EYFP-F
                     /note="color: black; sequence: CATCGACTTCAAGGAGGACGG"
     primer_bind     3253..3273
                     /label=EYFP-F
                     /note="color: black; sequence: CATCGACTTCAAGGAGGACGG"
     primer_bind     complement(3354..3376)
                     /label=EYFP-R
                     /note="color: black; sequence: GCGGATCTTGAAGTTCACCTTGA"
     misc_feature    complement(3369..3374)
                     /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"
     primer_bind     3485..3505
                     /label=EYFP-F2
                     /note="color: black; sequence: TCCAAGCTGAGCAAAGACCCC"
     misc_feature    3590..3617
                     /label=pEV542 (concatenated sequence 6)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3590..3593
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3614..3813
                     /label=pEV619 (concatenated sequence 7)
                     /note="Geneious type: Concatenated sequence"
                     /note="/Source"
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    3614..3617
                     /label=Ligation
                     /note="Geneious type: ligation"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    3614..3617
                     /label=CAAC
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
     polyA_signal    3618..3809
                     /label=SV40 polyA
                     /note="/created_by=User"
                     /note="/modified_by=elviravitu"
                     /note="color: #a6acb3; direction: RIGHT"
     protein_bind    3810..3813
                     /label=CGCT
                     /note="/created_by=User"
                     /note="color: #31849b; direction: RIGHT"
ORIGIN
        1 cgcttactga agagcggtct aatacccaat ctctcgtctt atccagatgt tttatacgcc
       61 ttaattaata gggataacag ggtaatatct tgccctgcac gaataccagc gaccccttgc
      121 ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg
      181 tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa
      241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
      661 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
      721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
      781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
      841 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
      901 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
      961 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1021 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1081 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1141 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     1201 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1261 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1321 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     1381 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     1441 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     1501 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     1561 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     1621 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     1681 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     1741 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     1801 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     1861 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     1921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     1981 tccgcgcacg aattggccag cgctgcatta ccctgttatc cctaaccggt gcgtttttat
     2041 gcttgtagta ttgtataatg tttttaagat ccgctcttca cgcggagttc tgcaattcat
     2101 gtgatctgcg cctgggagct cacggggaca gccccccccc aaagccccca gggatgtaat
     2161 tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct ccggtccggc
     2221 gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg gggaaggtgg
     2281 cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag acacctgggg
     2341 ggatacgggg aaaaagctag gctgaaagag gatcttagaa tgacacgcgc ctgggagctc
     2401 acggggacag ccccccccca aagcccccag ggatgtaatt acgtccctcc cccgctaggg
     2461 ggcagcagcg agccgcccgg ggctccgctc cggtccggcg ctccccccgc atccccgagc
     2521 cggcagcgtg cggggacagc ccgggcacgg ggaaggtggc acgggatcgc tttcctctga
     2581 acgcttctcg ctgctctttg agcctgcaga cacctggggg gatacgggga aaaagctgga
     2641 tcataactcg acgtgcatac tcggagtact gtcctccggc gattaacgga gtactgtcct
     2701 ccggcgatta acggagtact gtcctccggc gattaacgga gtactgtcct ccggcgatta
     2761 acggagttct gtcctccggg gcaccatatg ctctagaggg tatataatgg gggccaaatg
     2821 ctagtctact accagagctc atcgctagcg ctaccggtca ggtgccacca tggtgagcaa
     2881 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2941 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3001 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3061 cttcggctac ggcctgcagt gcttcgcccg ctaccccgac cacatgaagc agcacgactt
     3121 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3181 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3241 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3301 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3361 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3421 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     3481 ccagtccaag ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3541 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaag cttccactgg
     3601 attgtacaat taccaaccag acatgataag atacattgat gagtttggac aaaccacaac
     3661 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     3721 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     3781 ggttcagggg gaggtgtggg aggtttttt
//
