pYSJ29

LOCUS       pYSJ29             6920 bp DNA     circular SYN 26-OCT-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6920)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 5, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6920
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     897..955
                     /label=QL166 EWS-FL1-F
     primer_bind     913..955
                     /label=QL141-F
     primer_bind     914..955
                     /label=QL79 EWS-FLI1-F QL171-F
     primer_bind     914..954
                     /label=oYSJ79-EWS-F
     primer_bind     927..955
                     /label=QL168-F
     primer_bind     929..955
                     /label=QL107 EWS-F
     misc_feature    935..1726
                     /label=EWS
     primer_bind     935..954
                     /label=引物 1
     regulatory      935..938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1239..1260
                     /label=QL184 EWS-F
     primer_bind     complement(1343..1364)
                     /label=QL185 EWS-R
     primer_bind     1490..1512
                     /label=QL148
     primer_bind     complement(1573..1595)
                     /label=QL149
     primer_bind     1639..1661
                     /label=QL150
     primer_bind     1668..1689
                     /label=QL188 EF-F
     primer_bind     complement(1706..1726)
                     /label=QL 108 EWS-R
     primer_bind     complement(1706..1726)
                     /label=QL172-R
     primer_bind     complement(1706..1726)
                     /label=QL195 EWS-R
     primer_bind     complement(1706..1726)
                     /label=QL197 EWS-R(FEV)
     primer_bind     complement(1706..1726)
                     /label=QL199 EWS-R(ETV1)
     primer_bind     complement(1708..1746)
                     /label=oYSJ80-EWS-R
     primer_bind     complement(1708..1726)
                     /label=引物 2
     primer_bind     1710..1747
                     /label=oYSJ81-FEV-F
     misc_feature    1727..2443
                     /label=FEV
     primer_bind     2405..2416
                     /label=QL 108 EWS-R
     primer_bind     2405..2416
                     /label=QL172-R
     primer_bind     2405..2416
                     /label=QL195 EWS-R
     primer_bind     2405..2416
                     /label=QL197 EWS-R(FEV)
     primer_bind     2405..2416
                     /label=QL199 EWS-R(ETV1)
     primer_bind     complement(2421..2464)
                     /label=oYSJ82-FEV-R
     primer_bind     complement(2483..2502)
                     /label=QL 108 EWS-R
     polyA_signal    2520..2744
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2790..3218
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3232..3561
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3412..3547
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3628..4422
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4596..4717
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4766..4782)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4790..4806
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4814..4844)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4859..4880
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5168..5753)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5924..6784)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6785..6889)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccatggcg tccacggatt acagtaccta
      961 tagccaagct gcagcgcagc agggctacag tgcttacacc gcccagccca ctcaaggata
     1021 tgcacagacc acccaggcat atgggcaaca aagctatgga acctatggac agcccactga
     1081 tgtcagctat acccaggctc agaccactgc aacctatggg cagaccgcct atgcaacttc
     1141 ttatggacag cctcccactg gttatactac tccaactgcc ccccaggcat acagccagcc
     1201 tgtccagggg tatggcactg gtgcttatga taccaccact gctacagtca ccaccaccca
     1261 ggcctcctat gcagctcagt ctgcatatgg cactcagcct gcttatccag cctatgggca
     1321 gcagccagca gccactgcac ctacaagacc gcaggatgga aacaagccca ctgagactag
     1381 tcaacctcaa tctagcacag ggggttacaa ccagcccagc ctaggatatg gacagagtaa
     1441 ctacagttat ccccaggtac ctgggagcta ccccatgcag ccagtcactg cacctccatc
     1501 ctaccctcct accagctatt cctctacaca gccgactagt tatgatcaga gcagttactc
     1561 tcagcagaac acctatgggc aaccgagcag ctatggacag cagagtagct atggtcaaca
     1621 aagcagctat gggcagcagc ctcccactag ttacccaccc caaactggat cctacagcca
     1681 agctccaagt caatatagcc aacagagcag cagctacggg cagcagatga ggcagagtgg
     1741 agcttctcag cctctgctga tcaatatgta cctgcctgac cctgtgggtg atggactgtt
     1801 caaagacggc aagaatccat cttggggacc tctgtctcca gcagtgcaga agggatctgg
     1861 acagatccag ctgtggcagt tcttgctgga gttgctggct gacagagcca atgctggctg
     1921 tatcgcttgg gaaggtggac atggagagtt caagctgacc gatccagacg aagtggccag
     1981 aagatgggga gaacggaaga gcaagcctaa catgaattac gacaagctga gcagagcact
     2041 gaggtactac tacgacaaga acatcatgag caaggtgcac ggaaagaggt acgcctacag
     2101 gttcgacttc cagggcttgg ctcaggcttg tcagccacct ccagctcatg ctcatgctgc
     2161 tgccgcagca gctgccgccg ctgctgctgc tcaagacggt gctctgtaca agctgcctgc
     2221 tggattggct ccactgccat ttccaggtct gagcaagctg aacctgatgg ctgctagtgc
     2281 tggagtggct ccagctggct tcagctactg gcctggacct ggaccagctg ctacagcagc
     2341 cgctgcaaca gctgctctgt atccatctcc aagcctgcaa cctccacctg gacctttcgg
     2401 agctgtagct gctgcttctc atctgggtgg acactaccac tgagaattct gcagatatcc
     2461 agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     2521 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2581 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2641 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2701 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2761 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2821 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2881 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2941 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3001 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     3061 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     3121 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3181 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3241 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3361 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3421 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3481 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3541 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3601 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3661 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3721 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3781 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3841 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3901 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3961 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4021 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4081 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4141 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4201 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     4261 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     4321 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     4381 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     4441 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     4501 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4561 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4621 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4681 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4741 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4801 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4861 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4921 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4981 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5041 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     5101 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     5161 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     5221 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     5281 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     5341 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     5401 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     5461 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     5521 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5581 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5641 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5701 gcggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5761 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5821 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5881 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5941 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6001 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6061 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6121 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6181 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6241 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6301 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6361 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6421 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6481 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6541 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6601 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6661 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6721 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6781 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6841 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6901 gaaaagtgcc acctgacgtc
//