LOCUS       EW411_(UTRN-43-5bp_spac 7958 bp ds-DNA     circular     28-FEB-2025
DEFINITION  .
KEYWORDS    "accession:urn.local...1367881019035" "marker:5'TR, 3'TR, 6, 730 R,
            Xenopus globin 5'-UTR"
FEATURES             Location/Qualifiers
     enhancer        1..232
                     /label="core insul"
     primer          183..200
                     /label="EWXglobF NONSPECIFIC DONT USE"
                     /note="sequence: cttctcgctgctctttga"
     primer          224..239
                     /label="EWXglobF2"
                     /note="sequence: cggggaaaaggcctcc"
     primer_bind     246..246
                     /label="MluI-PGK-GF"
     misc_feature    246..246
                     /label="Gibson overlap"
     misc_feature    246..246
                     /label="Added MluI site"
     primer          248..286
                     /label="EWo173.1"
                     /note="sequence: aattgacgattgagtGGCAGGCCCGCAGCGGGGAGGAGG"
     primer          complement(252..291)
                     /label="EWo173.2"
                     /note="sequence: ctcctCCTCCTCCCCGCTGCGGGCCTGCCactcaatcgtc"
     misc_feature    263..286
                     /label="UTRN Promoter -43 Site"
     primer          287..321
                     /label="EWo174.3"
                     /note="sequence: aggagGAGCCGCCGAAGGAGCGAGCCTCTatcagt"
     misc_feature    292..315
                     /label="UTRN Promoter -14 Site"
     primer          complement(292..325)
                     /label="EWo174.4"
                     /note="sequence: ctagactgatAGAGGCTCGCTCCTTCGGCGGCTC"
     5'UTR           413..455
                     /label="Xenopus globin 5'-UTR"
     primer          complement(442..456)
                     /label="EWXglobR"
                     /note="sequence: gccaaagttgagcgt"
     misc_feature    491..491
                     /label="added SpeI site"
     misc_feature    492..497
                     /label="Added XhoI site"
     misc_feature    498..503
                     /label="Kozak"
     CDS             504..881
                     /label="H2B"
     CDS             504..893
                     /label="Translation 504-893"
                     /translation="MPEPAKSAPAPKKGSKKAVTKAQKKGGKKRKRSRKESYSIYVYKVLKQVHPDTGISSKAMGIMNSFVNDIFERIAGEASRLAHYNKRSTITSREIQTAVRLLLPGELAKHAVSEGTKAITKYTSAKDPRV"
     CDS             882..893
                     /label="linker"
     CDS             894..899
                     /label="linker"
     CDS             894..1613
                     /label="Translation 894-1613"
                     /translation="PVVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer          900..918
                     /label="EKo147.1"
                     /note="sequence: GTCTCAAAGGGCGAGGAAC"
     CDS             900..1613
                     /label="GFP"
     primer          complement(933..953)
                     /label="GFP5"
                     /note="sequence: CAATTCGACGAGGATCGGAAC"
     primer          complement(962..991)
                     /label="NK-gfp-seq-r"
                     /note="sequence: CCACTAACTGAGAATTTATGCCCATTAACG"
     primer          1048..1066
                     /label="pNSK084"
                     /note="sequence: CCGGAAAATTGCCAGTGCC"
     primer          complement(1156..1172)
                     /label="EKo224.2"
                     /note="sequence: ACCTTCAGGCATAGCGG"
     primer          1264..1284
                     /label="pPYK028"
                     /note="sequence: GGATCGAACTGAAGGGCATAG"
     primer          1541..1560
                     /label="GFP6"
                     /note="sequence: ACGGGATCACATGGTCCTTC"
     CDS             1614..1616
                     /label="Translation 1614-1616"
                     /translation="*"
     misc_feature    1622..1644
                     /label="MCS"
     primer          complement(1641..1658)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     polyA_signal    1645..1854
                     /label="BGH polyadenylation sequence"
     primer          complement(1720..1739)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          1831..1849
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     misc_feature    1854..1855
                     /label="GATC"
     misc_feature    1856..1863
                     /label="NotI"
     primer_bind     1864..1892
                     /label="5, 299 F"
     misc_feature    1898..1898
                     /label="ES003-2 C->A"
     misc_feature    1905..2213
                     /label="pSV40"
     misc_feature    2270..3295
                     /label="Hygro"
     primer_bind     2461..2477
                     /label="2, 563 F"
     primer          2501..2522
                     /label="EKo152.1"
                     /note="sequence: cttgacattggggaattcagcg"
     primer_bind     3033..3049
                     /label="3, 135 F"
     primer          3221..3239
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     misc_feature    complement(3275..3295)
                     /label="6, 730 R"
     misc_feature    3296..3301
                     /label="SalI"
     misc_feature    3302..3406
                     /label="WPRE"
     misc_feature    3412..3415
                     /label="SV40 pA(1)"
     misc_feature    3416..3419
                     /label="SV40 pA(1)"
     misc_feature    3417..3547
                     /label="SV40 pA"
     misc_feature    3420..3420
                     /label="SV40 pA(1)"
     misc_feature    3421..3421
                     /label="SV40 pA(1)"
     misc_feature    3422..3547
                     /label="SV40 pA(1)"
     primer          complement(3430..3455)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3454..3473)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     enhancer        complement(3796..4026)
                     /label="core insul(1) Geneious name: core insul(1)"
     repeat_region   4301..4335
                     /label="3'TR"
     misc_feature    4365..4365
                     /label="ES003-3 C->A"
     primer          complement(4826..4842)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(4840..4859)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer_bind     complement(4854..4873)
                     /label="pLac(F)"
     primer          5118..5137
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     primer          complement(5289..5308)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     misc_feature    complement(5987..6847)
                     /label="Amp"
     primer          complement(6112..6130)
                     /label="EKo152.2"
                     /note="sequence: tctcgcggtatcattgcag"
     primer          complement(7249..7267)
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     primer          7385..7400
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          7555..7571
                     /label="M13-40FOR Genewiz"
                     /note="sequence: GTTTTCCCAGTCACGAC"
     primer          7575..7590
                     /label="M13F Genewiz"
                     /note="sequence: GTAAAACGACGGCCAG"
     primer_bind     7659..7678
                     /label="BII(R)"
     repeat_region   7721..7733
                     /label="5'TR"
     misc_feature    7734..7764
                     /label="31bp spacer"
     repeat_region   7734..7764
                     /label="5'TR(1)"
     repeat_region   7765..7766
                     /label="5'TR"
     repeat_region   7768..7783
                     /label="5'TR"
ORIGIN
        1 gagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
       61 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tcccagagcc
      121 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
      181 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaggcctcca
      241 cggccacaat tgacgattga gtGGCAGGCC CGCAGCGGGG AGGAGGagga gGAGCCGCCG
      301 AAGGAGCGAG CCTCTatcag tctagagggt atataatgga tcccatcgcg tctcagcctc
      361 actttgagct cctccacacg aattccccga taccgtcgat tcaaggagct tgcttgttct
      421 ttttgcagaa gctcagaata aacgctcaac tttggcagat ctaccaagct tggtaccgag
      481 ctcggatcca actcgaggcc accatgccag agccagcgaa gtctgctccc gccccgaaaa
      541 agggctccaa gaaggcggtg actaaggcgc agaagaaagg cggcaagaag cgcaagcgca
      601 gccgcaagga gagctattcc atctatgtgt acaaggttct gaagcaggtc caccctgaca
      661 ccggcatttc gtccaaggcc atgggcatca tgaattcgtt tgtgaacgac attttcgagc
      721 gcatcgcagg tgaggcttcc cgcctggcgc attacaacaa gcgctcgacc atcacctcca
      781 gggagatcca gacggccgtg cgcctgctgc tgcctgggga gttggccaag cacgccgtgt
      841 ccgagggtac taaggccatc accaagtaca ccagcgctaa ggatccccgg gtaccggtcG
      901 TCTCAAAGGG CGAGGAACTT TTTACGGGCG TTGTTCCGAT CCTCGTCGAA TTGGACGGGG
      961 ACGTTAATGG GCATAAATTC TCAGTTAGTG GTGAGGGTGA GGGGGACGCG ACATATGGGA
     1021 AACTTACGCT GAAGTTCATA TGTACCACCG GAAAATTGCC aGTGCCGTGG CCTACTCTCG
     1081 TCACCACCCT GACATATGGT GTTCAGTGTT TTAGCCGGTA TCCAGACCAT ATGAAACAGC
     1141 ACGATTTTTT TAAATCCGCT ATGCCTGAAG GTTACGTACA GGAGCGAACG ATATTTTTTA
     1201 AAGACGACGG AAACTACAAA ACTAGAGCCG AGGTAAAATT CGAGGGTGAC ACTCTCGTGA
     1261 ACCGGATCGA ACTGAAGGGC ATAGATTTTA AGGAGGATGG CAATATACTG GGCCACAAGC
     1321 TCGAATACAA CTATAATTCT CATAATGTCT ATATCATGGC GGACAAACAG AAAAACGGCA
     1381 TCAAGGTAAA CTTTAAAATC CGACACAATA TTGAGGACGG CTCTGTCCAG CTTGCGGACC
     1441 ATTACCAACA GAACACGCCG ATAGGGGATG GTCCAGTACT TCTCCCTGAT AATCACTATT
     1501 TGTCTACTCA GTCAGCTCTT AGCAAGGACC CAAACGAAAA ACGGGATCAC ATGGTCCTTC
     1561 TTGAGTTCGT CACTGCGGCT GGGATAACAC TCGGGATGGA CGAACTGTAT AAGtagctag
     1621 cgaattcgaa tttaaatcgg cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1681 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     1741 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     1801 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggacgcggc
     1861 cgcaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     1921 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     1981 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2041 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2101 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2161 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2221 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtga tgaaaaagcc
     2281 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga
     2341 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg
     2401 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta
     2461 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag
     2521 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc
     2581 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc
     2641 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata
     2701 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac
     2761 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg
     2821 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt
     2881 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga
     2941 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca
     3001 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc
     3061 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga
     3121 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt
     3181 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt
     3241 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataggtcga
     3301 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     3361 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcgt taactaaact
     3421 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3481 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3541 atgtctggaa ttgactcaaa tgatgtcaat tagtctatca gaagctcatc tggtctccct
     3601 tccgggggac aagacatccc tgtttaatat ttaaacagca gtgttcccaa actgggttct
     3661 tatatccctt gctctggtca accaggttgc agggtttcct gtcctcacag gaacgaagtc
     3721 cctaaagaaa cagtggcagc caggtttagc cccggaattg actggattcc ttttttaggg
     3781 cccattggta tggctttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg
     3841 agaagcgttc agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca
     3901 cgctgccggc tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg
     3961 ctgctgcccc ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct
     4021 gtccctgata tctataacaa gaaaatatat atataataag ttatcacgta agtagaacat
     4081 gaaataacaa tataattatc gtatgagtta aatcttaaaa gtcacgtaaa agataatcat
     4141 gcgtcatttt gactcacgcg gtcgttatag ttcaaaatca gtgacactta ccgcattgac
     4201 aagcacgcct cacgggagct ccaagcggcg actgagatgt cctaaatgca cagcgacgga
     4261 ttcgcgctat ttagaaagag agagcaatat ttcaagaatg catgcgtcaa ttttacgcag
     4321 actatctttc tagggttaat ctagctgcat caggatcata tcgtcgggtc ttttttccgg
     4381 ctcagtcatc gcccaagctg gcgctatctg ggcatcgggg aggaagaagc ccgtgccttt
     4441 tcccgcgagg ttgaagcggc atggaaagag tttgccgagg atgactgctg ctgcattgac
     4501 gttgagcgaa aacgcacgtt taccatgatg attcgggaag gtgtggccat gcacgccttt
     4561 aacggtgaac tgttcgttca ggccacctgg gataccagtt cgtcgcggct tttccggaca
     4621 cagttccgga tggtcagccc gaagcgcatc agcaacccga acaataccgg cgacagccgg
     4681 aactgccgtg ccggtgtgca gattaatgac agcggtgcgg cgctgggata ttacgtcagc
     4741 gaggacgggt atcctggctg gatgccgcag aaatggacat ggataccccg tgagttaccc
     4801 ggcgggcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4861 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4921 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4981 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5041 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5101 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5161 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5221 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5281 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5341 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5401 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5461 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5521 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5581 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5641 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5701 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5761 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5821 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5881 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5941 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6001 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6061 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     6121 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     6181 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     6241 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     6301 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     6361 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     6421 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     6481 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     6541 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6601 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6661 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6721 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6781 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6841 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6901 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6961 cccgaaaagt gccacctaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     7021 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     7081 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     7141 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     7201 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     7261 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     7321 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     7381 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     7441 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     7501 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     7561 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcc tcgttcattc acgtttttga
     7621 acccgtggag gacgggcaga ctcgcggtgc aaatgtgttt tacagcgtga tggagcagat
     7681 gaagatgctc gacacgctgc agaacacgca gctagattaa ccctagaaag ataatcatat
     7741 tgtgacgtac gttaaagata atcatgtgta aaattgacgc atgtgtttta tcggtctgta
     7801 tatcgaggtt tatttattaa tttgaataga tattaagttt tattatattt acacttacat
     7861 actaataata aattcaacaa acaatttatt tatgtttatt tatttattaa aaaaaacaaa
     7921 aactcaaaat ttcttctata aagtaacaaa acttttat
//