pLYL85

LOCUS       pLYL85               6394 bp DNA     circular SYN 13-JAN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6394)
  AUTHORS   iGEM CPU-CHINA 2020
  TITLE     Direct Submission
  JOURNAL   Exported Jul 18, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6394
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     166..191
                     /label=182-WKG-596-F
     misc_feature    234..245
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    248..259
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    262..273
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    276..287
                     /label=VanO
                     /label=nonstandard type:  operator
     misc_feature    290..301
                     /label=VanO
                     /label=nonstandard type:  operator
     promoter        324..355
                     /label=P_min
     misc_feature    357..361
                     /label=NheI
     misc_feature    372
                     /label=New Feature
     misc_feature    372
                     /label=New Feature
     regulatory      378..387
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             386
                     /codon_start=1
                     /label=5xGS Linker
                     /translation=""
     CDS             387..449
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             450..524
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     CDS             525..587
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             588..662
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     CDS             669..731
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             732..806
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     CDS             807..869
                     /codon_start=1
                     /label=ANR
                     /translation="GELDELVYLLDGPGYDPIHSD"
     CDS             870..944
                     /codon_start=1
                     /label=5xGS Linker
                     /translation="GSGTGSGTGSGTGTTSGTGTGGSTG"
     misc_feature    951..1892
                     /label=NSP3
     misc_feature    951..974
                     /label=NSP3
     primer_bind     complement(962..981)
                     /label=179-WKG-592-R
     misc_feature    1915..1920
                     /label=EcoRI
     misc_feature    1954..1959
                     /label=New Feature
     misc_feature    1954..1959
                     /label=New Feature
     polyA_signal    1991..2215
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2261..2689
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2703..3032
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3099..3893
                     /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    4067..4188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4233..4253)
                     /label=M13-rev
     primer_bind     complement(4237..4253)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(4259..4281)
                     /label=LacO
     protein_bind    4261..4277
                     /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(4285..4315)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(4286..4315)
                     /label=lac
     protein_bind    4330..4351
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    4457..5397
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(5398..6258)
                     /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(6259..6363)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6300..6328
                     /label=231
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 cgtattggat
      241 ccaatgcatt ggatccaatg gattggatcc aatcgattgg atccaattga ttggatccaa
      301 tttaattaag cagctcctgc aggtagaggg tatataatgg aagctcgact tccaggctag
      361 cgtttaaact taagcttgcc accatggggg agctcgacga gttggtttat ctcttagacg
      421 gacccggata tgaccccatt cattcagatg gctccggaac gggcagcggg accggaagtg
      481 gcaccggaac cacttccgga accggtactg gcgggagtac cggcggtgag ctagacgaac
      541 tcgtgtacct gttggatggc cccggttacg acccaatcca cagcgatgga agtggaactg
      601 ggtctgggac tgggtccggc accgggacta cttcaggcac cggcactggg ggctccactg
      661 gccataccgg ggagctcgac gagttggttt atctcttaga cggacccgga tatgacccca
      721 ttcattcaga tggctccgga acgggcagcg ggaccggaag tggcaccgga accacttccg
      781 gaaccggtac tggcgggagt accggcggtg agctagacga actcgtgtac ctgttggatg
      841 gccccggtta cgacccaatc cacagcgatg gaagtggaac tgggtctggg actgggtccg
      901 gcaccgggac tacttcaggc accggcactg ggggctccac tggccatatg atgctcaaga
      961 tggagtctac acagcagatg gctagttcga ttattaacac ttcttttgaa gccgcagtcg
     1021 ttgctgccac ttcaactctt gaacttatgg gcattcaata tgattataat gaagtttata
     1081 ctagagttaa gagcaaattt gattacgtaa tggatgattc aggtgttaaa aacaatctgt
     1141 tgggtaaagc ggcaactatt gatcaggcac tgaatggaaa atttggttca gcagcaagaa
     1201 atagaaattg gatggctgat actagaacta cagctagact agatgaagat gtgaacaaac
     1261 ttagaatgat gctatcatct aaaggaattg atcaaaaaat gagagtactt aatgcatgtt
     1321 tcaatgtgaa gcgagttcca ggtaaatcat catcaataat taaatgtact agattaatga
     1381 gagataagat tgaacgtggt gaagtcgaag tggatgattc gtttgttgaa gaaaaaatgg
     1441 aagttgacac aattgattgg aaatctagat atgaacaatt ggaaaaacga tttgaatcgc
     1501 tgaaacaaag ggtgaatgaa aaatatacaa gttgggtgca aaaagcaaaa aaggtaaatg
     1561 aaaatatgta ctctctccaa aacgttatat cacaacagca gagccagatt gctgatttac
     1621 aaaattactg taataaattg gaagttgatc tacaaaataa aattagttca cttgtgtctt
     1681 cagtcgaatg gtacctgaaa tcaatggaat tgcctgatga aattaagaca gatattgaac
     1741 aacaactaaa ttccattgat gtaatcaatc caattaatgc tatagatgat tttgaatcac
     1801 tgatcagaaa tattatactc gactatgaca gaatattttt aatgtttaaa ggattaatga
     1861 ggcaatgcaa ctatgaatat acgtatgaat aggatccact agtccagtgt ggtggaattc
     1921 tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta aacccgctga
     1981 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2041 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2101 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2161 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2221 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2281 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2341 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2401 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2461 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2521 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2581 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2641 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2701 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2761 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2821 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2881 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2941 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3001 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3061 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3121 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3181 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3241 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3301 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3361 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3421 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3481 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3541 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3601 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3661 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3721 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3781 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3841 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3901 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3961 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4021 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4201 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4261 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4321 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4381 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4441 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4501 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4561 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4621 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4681 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4741 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4801 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4861 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4921 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4981 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5041 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5101 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5161 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5221 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5281 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5341 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     5401 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     5461 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     5521 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     5581 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     5641 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5701 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5761 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5821 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5881 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5941 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     6001 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     6061 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     6121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     6181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     6241 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     6301 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     6361 gcgcacattt ccccgaaaag tgccacctga cgtc
//