pSL549

LOCUS       pSL549              6933 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 6933)
  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..6933
                     /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     643..668
                     /label=185-WKG-602-CMV-F
     primer_bind     863..882
                     /label=T7
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    895..900
                     /label=NheI
     misc_feature    911
                     /label=New Feature
     misc_feature    911
                     /label=New Feature
     regulatory      917..926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             925
                     /codon_start=1
                     /label=5xGS Linker
                     /translation=""
     CDS             926..988
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             989..1063
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     CDS             1064..1126
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             1127..1201
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     CDS             1208..1270
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             1271..1345
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     CDS             1346..1408
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             1409..1483
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     misc_feature    1490..2431
                     /label=NSP3
     misc_feature    1490..1513
                     /label=NSP3
     primer_bind     complement(1501..1520)
                     /label=179-WKG-592-R
     misc_feature    2454..2459
                     /label=EcoRI
     misc_feature    2493..2498
                     /label=New Feature
     misc_feature    2493..2498
                     /label=New Feature
     polyA_signal    2530..2754
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2800..3228
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     3099..3118
                     /label=189-WKG-612-F
     promoter        3242..3571
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     misc_feature    3422..3499
                     /label=SV40 ORI
     CDS             3638..4432
                     /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"
     primer_bind     complement(3682..3709)
                     /label=188-WKG-610-R
     polyA_signal    4606..4727
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4772..4792)
                     /label=M13-rev
     primer_bind     complement(4776..4792)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(4798..4820)
                     /label=LacO
     protein_bind    4800..4816
                     /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(4824..4854)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(4825..4854)
                     /label=lac
     protein_bind    4869..4890
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    4996..5936
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(5937..6797)
                     /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(6798..6902)
                     /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 aagcttgcca ccatggggga gctcgacgag ttggtttatc tcttagacgg
      961 acccggatat gaccccattc attcagatgg ctccggaacg ggcagcggga ccggaagtgg
     1021 caccggaacc acttccggaa ccggtactgg cgggagtacc ggcggtgagc tagacgaact
     1081 cgtgtacctg ttggatggcc ccggttacga cccaatccac agcgatggaa gtggaactgg
     1141 gtctgggact gggtccggca ccgggactac ttcaggcacc ggcactgggg gctccactgg
     1201 ccataccggg gagctcgacg agttggttta tctcttagac ggacccggat atgaccccat
     1261 tcattcagat ggctccggaa cgggcagcgg gaccggaagt ggcaccggaa ccacttccgg
     1321 aaccggtact ggcgggagta ccggcggtga gctagacgaa ctcgtgtacc tgttggatgg
     1381 ccccggttac gacccaatcc acagcgatgg aagtggaact gggtctggga ctgggtccgg
     1441 caccgggact acttcaggca ccggcactgg gggctccact ggccatatga tgctcaagat
     1501 ggagtctaca cagcagatgg ctagttcgat tattaacact tcttttgaag ccgcagtcgt
     1561 tgctgccact tcaactcttg aacttatggg cattcaatat gattataatg aagtttatac
     1621 tagagttaag agcaaatttg attacgtaat ggatgattca ggtgttaaaa acaatctgtt
     1681 gggtaaagcg gcaactattg atcaggcact gaatggaaaa tttggttcag cagcaagaaa
     1741 tagaaattgg atggctgata ctagaactac agctagacta gatgaagatg tgaacaaact
     1801 tagaatgatg ctatcatcta aaggaattga tcaaaaaatg agagtactta atgcatgttt
     1861 caatgtgaag cgagttccag gtaaatcatc atcaataatt aaatgtacta gattaatgag
     1921 agataagatt gaacgtggtg aagtcgaagt ggatgattcg tttgttgaag aaaaaatgga
     1981 agttgacaca attgattgga aatctagata tgaacaattg gaaaaacgat ttgaatcgct
     2041 gaaacaaagg gtgaatgaaa aatatacaag ttgggtgcaa aaagcaaaaa aggtaaatga
     2101 aaatatgtac tctctccaaa acgttatatc acaacagcag agccagattg ctgatttaca
     2161 aaattactgt aataaattgg aagttgatct acaaaataaa attagttcac ttgtgtcttc
     2221 agtcgaatgg tacctgaaat caatggaatt gcctgatgaa attaagacag atattgaaca
     2281 acaactaaat tccattgatg taatcaatcc aattaatgct atagatgatt ttgaatcact
     2341 gatcagaaat attatactcg actatgacag aatattttta atgtttaaag gattaatgag
     2401 gcaatgcaac tatgaatata cgtatgaata ggatccacta gtccagtgtg gtggaattct
     2461 gcagatatcc agcacagtgg cggccgctcg agtctagagg gcccgtttaa acccgctgat
     2521 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     2581 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     2641 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     2701 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     2761 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     2821 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     2881 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     2941 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3001 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3061 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3121 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3181 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     3241 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3301 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3361 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3421 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3481 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3541 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3601 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3661 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3721 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3781 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3841 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3901 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3961 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4021 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4081 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4141 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4201 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4261 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4321 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4381 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4441 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4501 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4561 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4621 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4681 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4741 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4801 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4861 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4921 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4981 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5041 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5101 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5161 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5221 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5281 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5341 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5401 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5461 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5521 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5581 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5641 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5701 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5761 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5821 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5881 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5941 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     6001 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6061 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6121 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6181 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6241 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6301 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6361 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6421 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6481 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6541 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6601 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6661 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6721 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6781 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6841 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6901 cgcacatttc cccgaaaagt gccacctgac gtc
//