pLYL87

LOCUS       pLYL87              8100 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 8100)
  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..8100
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     166..191
                     /label=182-WKG-596-F
     misc_feature    232
                     /label=CMV
     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    362
                     /label=CMV
     primer_bind     406..425
                     /label=T7
     promoter        406..424
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      443..452
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             452..1150
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    1167..1185
                     /label=linker
     CDS             1187..1885
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    1902..1920
                     /label=linker
     CDS             1922..2620
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    2637..2656
                     /label=linker
     misc_feature    2657..3598
                     /label=NSP3
     misc_feature    2657..2670
                     /label=NSP3
     primer_bind     complement(2668..2687)
                     /label=179-WKG-592-R
     polyA_signal    3697..3921
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3967..4395
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4409..4738
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4805..5599
                     /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    5773..5894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5939..5959)
                     /label=M13-rev
     primer_bind     complement(5943..5959)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(5965..5987)
                     /label=LacO
     protein_bind    5967..5983
                     /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(5991..6021)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(5992..6021)
                     /label=lac
     protein_bind    6036..6057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6163..7103
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(7104..7964)
                     /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(7965..8069)
                     /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 cgtattggat
      241 ccaatgcatt ggatccaatg gattggatcc aatcgattgg atccaattga ttggatccaa
      301 tttaattaag cagctcctgc aggtagaggg tatataatgg aagctcgact tccagggagc
      361 tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta
      421 tagggagacc caagctggct aggccaccat ggacatcgag aagctgtgca agaaggcgga
      481 agaggaggcc aaggaggccc aggagaaggc cgacgagctg agacaacgac accccgattc
      541 gcaggcggcg gaagacgcgg aggatctcgc aaatctggct gtcgccgccg tgctgaccgc
      601 ttgtctgctg gcccaggagc atcccaacgc cgatatcgct aagctgtgca tcaaggccgc
      661 gagcgaagct gcggaggccg cgtcaaaagc cgccgagctg gcccaacggc acccagacag
      721 tcaagcagcc agagatgcta tcaaactcgc ctctcaagcc gcccgcgccg tgatcctggc
      781 catcatgctc gccgcagaga atcccaacgc tgatattgca aagctatgta ttaaggcggc
      841 cagcgaagcc gcagaggctg cctcgaaagc tgcggaactt gcacagcggc accccgactc
      901 ccaggccgcc cgagatgcaa ttaaactggc cagtcaggcc gctgaggcgg tggagagagc
      961 catctggctg gccgccgaga accccaatgc cgatatcgcc aaaaaatgca tcaaagcggc
     1021 ctccgaagca gccgaggagg cctctaaagc tgccgaagag gctcaaagac acccagacag
     1081 ccagaaggca cgcgacgaga tcaaggaggc ctctcagaag gccgaggagg tgaaggagag
     1141 atgcaagagc gccagcggaa gtggcggagg aggcgacgtc gctagtgaca tcgagaagct
     1201 gtgcaagaag gcggaagagg aggccaagga ggcccaggag aaggccgacg agctgagaca
     1261 acgacacccc gattcgcagg cggcggaaga cgcggaggat ctcgcaaatc tggctgtcgc
     1321 cgccgtgctg accgcttgtc tgctggccca ggagcatccc aacgccgata tcgctaagct
     1381 gtgcatcaag gccgcgagcg aagctgcgga ggccgcgtca aaagccgccg agctggccca
     1441 acggcaccca gacagtcaag cagccagaga tgctatcaaa ctcgcctctc aagccgcccg
     1501 cgccgtgatc ctggccatca tgctcgccgc agagaatccc aacgctgata ttgcaaagct
     1561 atgtattaag gcggccagcg aagccgcaga ggctgcctcg aaagctgcgg aacttgcaca
     1621 gcggcacccc gactcccagg ccgcccgaga tgcaattaaa ctggccagtc aggccgctga
     1681 ggcggtggag agagccatct ggctggccgc cgagaacccc aatgccgata tcgccaaaaa
     1741 atgcatcaaa gcggcctccg aagcagccga ggaggcctct aaagctgccg aagaggctca
     1801 aagacaccca gacagccaga aggcacgcga cgagatcaag gaggcctctc agaaggccga
     1861 ggaggtgaag gagagatgca agagcgccag cggaagtggc ggaggaggcg acgtcgctag
     1921 tgacatcgag aagctgtgca agaaggcgga agaggaggcc aaggaggccc aggagaaggc
     1981 cgacgagctg agacaacgac accccgattc gcaggcggcg gaagacgcgg aggatctcgc
     2041 aaatctggct gtcgccgccg tgctgaccgc ttgtctgctg gcccaggagc atcccaacgc
     2101 cgatatcgct aagctgtgca tcaaggccgc gagcgaagct gcggaggccg cgtcaaaagc
     2161 cgccgagctg gcccaacggc acccagacag tcaagcagcc agagatgcta tcaaactcgc
     2221 ctctcaagcc gcccgcgccg tgatcctggc catcatgctc gccgcagaga atcccaacgc
     2281 tgatattgca aagctatgta ttaaggcggc cagcgaagcc gcagaggctg cctcgaaagc
     2341 tgcggaactt gcacagcggc accccgactc ccaggccgcc cgagatgcaa ttaaactggc
     2401 cagtcaggcc gctgaggcgg tggagagagc catctggctg gccgccgaga accccaatgc
     2461 cgatatcgcc aaaaaatgca tcaaagcggc ctccgaagca gccgaggagg cctctaaagc
     2521 tgccgaagag gctcaaagac acccagacag ccagaaggca cgcgacgaga tcaaggaggc
     2581 ctctcagaag gccgaggagg tgaaggagag atgcaagagc gccagcggaa gtggcggagg
     2641 aggcgacgtc gctagcatgc tcaagatgga gtctacacag cagatggcta gttcgattat
     2701 taacacttct tttgaagccg cagtcgttgc tgccacttca actcttgaac ttatgggcat
     2761 tcaatatgat tataatgaag tttatactag agttaagagc aaatttgatt acgtaatgga
     2821 tgattcaggt gttaaaaaca atctgttggg taaagcggca actattgatc aggcactgaa
     2881 tggaaaattt ggttcagcag caagaaatag aaattggatg gctgatacta gaactacagc
     2941 tagactagat gaagatgtga acaaacttag aatgatgcta tcatctaaag gaattgatca
     3001 aaaaatgaga gtacttaatg catgtttcaa tgtgaagcga gttccaggta aatcatcatc
     3061 aataattaaa tgtactagat taatgagaga taagattgaa cgtggtgaag tcgaagtgga
     3121 tgattcgttt gttgaagaaa aaatggaagt tgacacaatt gattggaaat ctagatatga
     3181 acaattggaa aaacgatttg aatcgctgaa acaaagggtg aatgaaaaat atacaagttg
     3241 ggtgcaaaaa gcaaaaaagg taaatgaaaa tatgtactct ctccaaaacg ttatatcaca
     3301 acagcagagc cagattgctg atttacaaaa ttactgtaat aaattggaag ttgatctaca
     3361 aaataaaatt agttcacttg tgtcttcagt cgaatggtac ctgaaatcaa tggaattgcc
     3421 tgatgaaatt aagacagata ttgaacaaca actaaattcc attgatgtaa tcaatccaat
     3481 taatgctata gatgattttg aatcactgat cagaaatatt atactcgact atgacagaat
     3541 atttttaatg tttaaaggat taatgaggca atgcaactat gaatatacgt atgaatagga
     3601 tccactagtc cagtgtggtg gaattctgca gatatccagc acagtggcgg ccgctcgagt
     3661 ctagagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat
     3721 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3781 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3841 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3901 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     3961 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4021 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4081 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4141 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4201 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4261 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     4321 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4381 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4441 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4501 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4561 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4621 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     4681 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     4741 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     4801 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4861 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4921 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4981 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5041 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5101 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5161 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5221 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     5281 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     5341 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     5401 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     5461 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     5521 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     5581 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     5641 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5701 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5761 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5821 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5881 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     5941 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6001 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6061 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6121 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6181 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6241 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6301 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6361 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6421 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6481 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6541 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6601 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6661 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6721 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6781 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6841 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6901 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6961 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7021 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7081 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7141 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7201 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7261 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7321 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7381 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7441 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7501 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7561 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7621 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7681 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7741 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7801 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7861 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7921 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7981 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8041 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//