pSL548

LOCUS       pSL548             7673 bp DNA     circular SYN 14-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7673)
  AUTHORS   Westlake University
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7673
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     primer_bind     863..882
                     /label=T7
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      906..914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             914..1300
                     /codon_start=1
                     /product="bacteriophage MS2 coat protein"
                     /label=MS2-N55K
                     /note="binds with high affinity to a specific stem-loop 
                     structure in the viral RNA (Lim et al., 1994)"
                     /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT
                     CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI
                     VKAMQGLLKDGNPIPSAIAANSGIY"
     primer_bind     1231..1257
                     /label=177-WKG-589-F
     misc_feature    1301..1336
                     /label=linker
     misc_feature    1301..1321
                     /label=linker
     CDS             1337..1924
                     /codon_start=1
                     /label=NS3a H1
                     /translation="KKKGSVVIVGRINLSGDTAYSQQTRGLEGCQETSQTGRDKNQVEG
                     EVQVVSTATQSFLATSINGVLWTVYHGAGTRTIASPKGPVTQMYTNVDKDLVGWQAPQG
                     SRSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGH
                     AVGIFRAAVSTRGVAKAVDFIPVESLETTMRSP"
     misc_feature    1931..1966
                     /label=linker
     misc_feature    1931..1951
                     /label=linker
     CDS             1967..2554
                     /codon_start=1
                     /label=NS3a H1
                     /translation="KKKGSVVIVGRINLSGDTAYSQQTRGLEGCQETSQTGRDKNQVEG
                     EVQVVSTATQSFLATSINGVLWTVYHGAGTRTIASPKGPVTQMYTNVDKDLVGWQAPQG
                     SRSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGH
                     AVGIFRAAVSTRGVAKAVDFIPVESLETTMRSP"
     misc_feature    2561..2596
                     /label=linker
     misc_feature    2561..2581
                     /label=linker
     CDS             2597..3184
                     /codon_start=1
                     /label=NS3a H1
                     /translation="KKKGSVVIVGRINLSGDTAYSQQTRGLEGCQETSQTGRDKNQVEG
                     EVQVVSTATQSFLATSINGVLWTVYHGAGTRTIASPKGPVTQMYTNVDKDLVGWQAPQG
                     SRSLTPCTCGSSDLYLVTRHADVIPVRRRGDSRGSLLSPRPISYLKGSSGGPLLCPAGH
                     AVGIFRAAVSTRGVAKAVDFIPVESLETTMRSP"
     primer_bind     complement(3185..3213)
                     /label=205-WKG-647-R
     polyA_signal    3270..3494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3540..3968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3554..3578)
                     /label=184-WKG-599-R
     primer_bind     3839..3858
                     /label=189-WKG-612-F
     promoter        3982..4311
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4378..5172
                     /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    5346..5467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5512..5532)
                     /label=M13-rev
     primer_bind     complement(5516..5532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(5538..5560)
                     /label=LacO
     protein_bind    5540..5556
                     /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(5564..5594)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(5565..5594)
                     /label=lac
     protein_bind    5609..5630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    5736..6676
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(6677..7537)
                     /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(7538..7642)
                     /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 gctggctagg
      901 aattcccacc atggcttcaa actttactca gttcgtgctc gtggacaatg gtgggacagg
      961 ggatgtgaca gtggctcctt ctaatttcgc taatggggtg gcagagtgga tcagctccaa
     1021 ctcacggagc caggcctaca aggtgacatg cagcgtcagg cagtctagtg cccagaagag
     1081 aaagtatacc atcaaggtgg aggtccccaa agtggctacc cagacagtgg gcggagtcga
     1141 actgcctgtc gccgcttgga ggtcctacct gaacatggag ctcactatcc caattttcgc
     1201 taccaattct gactgtgaac tcatcgtgaa ggcaatgcag gggctcctca aagacggtaa
     1261 tcctatccct tccgccatcg ccgctaactc aggtatctac gccagcggaa gtggcggagg
     1321 aggcgacgtc ggatccaaga agaagggcag cgtggtgatc gtgggcagaa tcaacctgag
     1381 cggcgacacc gcctacagcc agcagaccag aggcctggag ggctgccagg agaccagcca
     1441 gaccggcaga gacaagaacc aggtggaggg cgaggtgcag gtggtgagca ccgccaccca
     1501 gagcttcctg gccaccagca tcaacggcgt gctgtggacc gtgtaccacg gcgccggcac
     1561 cagaaccatc gccagcccca agggccccgt gacccagatg tacacgaacg tggacaagga
     1621 cttagtgggc tggcaggccc cccaaggcag cagaagcctg accccctgca cctgcggcag
     1681 cagcgacctg tacctggtga ccagacacgc cgacgtgatc cccgtgagaa gaagaggcga
     1741 cagcagaggc agcctgctga gccccagacc catcagctat ctgaaaggct caagcggcgg
     1801 ccccctgttg tgtcccgccg gccatgccgt gggcatcttc agagccgccg tgagcaccag
     1861 aggcgtggcc aaggccgtgg acttcatccc cgtggagagc ctggagacca ccatgagaag
     1921 ccccgctagt gccagcggaa gtggcggagg aggcgacgtc ggatccaaga agaagggcag
     1981 cgtggtgatc gtgggcagaa tcaacctgag cggcgacacc gcctacagcc agcagaccag
     2041 aggcctggag ggctgccagg agaccagcca gaccggcaga gacaagaacc aggtggaggg
     2101 cgaggtgcag gtggtgagca ccgccaccca gagcttcctg gccaccagca tcaacggcgt
     2161 gctgtggacc gtgtaccacg gcgccggcac cagaaccatc gccagcccca agggccccgt
     2221 gacccagatg tacacgaacg tggacaagga cttagtgggc tggcaggccc cccaaggcag
     2281 cagaagcctg accccctgca cctgcggcag cagcgacctg tacctggtga ccagacacgc
     2341 cgacgtgatc cccgtgagaa gaagaggcga cagcagaggc agcctgctga gccccagacc
     2401 catcagctat ctgaaaggct caagcggcgg ccccctgttg tgtcccgccg gccatgccgt
     2461 gggcatcttc agagccgccg tgagcaccag aggcgtggcc aaggccgtgg acttcatccc
     2521 cgtggagagc ctggagacca ccatgagaag ccccgctagt gccagcggaa gtggcggagg
     2581 aggcgacgtc ggatccaaga agaagggcag cgtggtgatc gtgggcagaa tcaacctgag
     2641 cggcgacacc gcctacagcc agcagaccag aggcctggag ggctgccagg agaccagcca
     2701 gaccggcaga gacaagaacc aggtggaggg cgaggtgcag gtggtgagca ccgccaccca
     2761 gagcttcctg gccaccagca tcaacggcgt gctgtggacc gtgtaccacg gcgccggcac
     2821 cagaaccatc gccagcccca agggccccgt gacccagatg tacacgaacg tggacaagga
     2881 cttagtgggc tggcaggccc cccaaggcag cagaagcctg accccctgca cctgcggcag
     2941 cagcgacctg tacctggtga ccagacacgc cgacgtgatc cccgtgagaa gaagaggcga
     3001 cagcagaggc agcctgctga gccccagacc catcagctat ctgaaaggct caagcggcgg
     3061 ccccctgttg tgtcccgccg gccatgccgt gggcatcttc agagccgccg tgagcaccag
     3121 aggcgtggcc aaggccgtgg acttcatccc cgtggagagc ctggagacca ccatgagaag
     3181 ccccgctagc tagaaattct gcagatatcc agcacagtgg cggccgctcg agtctagagg
     3241 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3301 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3361 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     3421 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     3481 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     3541 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     3601 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     3661 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     3721 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     3781 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     3841 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     3901 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3961 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     4021 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     4141 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     4201 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     4261 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     4321 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     4381 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4441 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4501 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     4561 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4621 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4681 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4741 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4801 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4861 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     4921 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4981 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5041 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5101 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5161 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5221 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5281 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5341 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5401 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5461 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     5521 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     5581 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     5641 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     5701 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     5761 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     5821 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     5881 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     5941 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     6001 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     6061 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     6121 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     6181 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6241 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     6301 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     6361 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     6421 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     6481 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     6541 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     6601 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     6661 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     6721 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6781 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6841 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6901 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6961 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     7021 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     7081 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     7141 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     7201 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     7261 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     7321 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     7381 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     7441 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     7501 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     7561 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     7621 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//