pLYL62

LOCUS       pLYL62            8444 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 8444)
  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..8444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    232
                     /label=CMV
     primer_bind     234..250
                     /label=oMH18
     misc_feature    251..404
                     /label=Transcription blocker
                     /label=Transcription\blocker
     misc_feature    438..524
                     /label=CRE reg_Regulatory_Seq
     misc_feature    449..456
                     /label=CRE
     misc_feature    485..492
                     /label=CRE*
     misc_feature    507..514
                     /label=CRE
     promoter        531..677
                     /label=P_TAL
     primer_bind     634..661
                     /label=180-WKG-592-F
     misc_feature    706
                     /label=CMV
     primer_bind     750..769
                     /label=T7
     promoter        750..768
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      787..796
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             796..1494
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    1511..1529
                     /label=linker
     CDS             1531..2229
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    2246..2264
                     /label=linker
     CDS             2266..2964
                     /codon_start=1
                     /label=GNCR1-N
                     /translation="DIEKLCKKAEEEAKEAQEKADELRQRHPDSQAAEDAEDLANLAVA
                     AVLTACLLAQEHPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQAA
                     RAVILAIMLAAENPNADIAKLCIKAASEAAEAASKAAELAQRHPDSQAARDAIKLASQA
                     AEAVERAIWLAAENPNADIAKKCIKAASEAAEEASKAAEEAQRHPDSQKARDEIKEASQ
                     KAEEVKERCKS"
     misc_feature    2981..3000
                     /label=linker
     misc_feature    3001..3942
                     /label=NSP3
     misc_feature    3001..3014
                     /label=NSP3
     primer_bind     complement(3012..3031)
                     /label=179-WKG-592-R
     primer_bind     complement(3991..4000)
                     /label=oMH18
     terminator      4038..4265
                     /label=bGH PA
     rep_origin      4311..4739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4753..5082
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     misc_feature    4933..5010
                     /label=SV40 ORI
     CDS             5149..5943
                     /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    6117..6238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6283..6303)
                     /label=M13-rev
     primer_bind     complement(6287..6303)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_binding    complement(6309..6331)
                     /label=LacO
     protein_bind    6311..6327
                     /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(6335..6365)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        complement(6336..6365)
                     /label=lac
     protein_bind    6380..6401
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    6507..7447
                     /label=ColE1
                     /note="ColE1"
     CDS             complement(7448..8308)
                     /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(8309..8413)
                     /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 cgtgtacttg
      241 gagcggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
      301 atcgatagta ctaacatacg ctctccatca aaacaaaacg aaacaaaaca aactagcaaa
      361 ataggctgtc cccagtgcaa gtgcaggtgc cagaacattt ctctatcgat aggtaccgag
      421 ctcttacgcg tgctagcgca ccagacagtg acgtcagctg ccagatccca tggccgtcat
      481 actgtgacgt ctttcagaca ccccattgac gtcaatggga gaacagatct gccgccccga
      541 ctgcatctgc gtgttcgaat tcgccaatga caagacgctg ggcggggttt gtgtcatcat
      601 agaactaaag acatgcaaat atatttcttc cggggacacc gccagcaaac gcgagcaacg
      661 ggccacgggg atgaagcaga agcttcgaat cgcgaattcg gagctctctg gctaactaga
      721 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
      781 ggctaggcca ccatggacat cgagaagctg tgcaagaagg cggaagagga ggccaaggag
      841 gcccaggaga aggccgacga gctgagacaa cgacaccccg attcgcaggc ggcggaagac
      901 gcggaggatc tcgcaaatct ggctgtcgcc gccgtgctga ccgcttgtct gctggcccag
      961 gagcatccca acgccgatat cgctaagctg tgcatcaagg ccgcgagcga agctgcggag
     1021 gccgcgtcaa aagccgccga gctggcccaa cggcacccag acagtcaagc agccagagat
     1081 gctatcaaac tcgcctctca agccgcccgc gccgtgatcc tggccatcat gctcgccgca
     1141 gagaatccca acgctgatat tgcaaagcta tgtattaagg cggccagcga agccgcagag
     1201 gctgcctcga aagctgcgga acttgcacag cggcaccccg actcccaggc cgcccgagat
     1261 gcaattaaac tggccagtca ggccgctgag gcggtggaga gagccatctg gctggccgcc
     1321 gagaacccca atgccgatat cgccaaaaaa tgcatcaaag cggcctccga agcagccgag
     1381 gaggcctcta aagctgccga agaggctcaa agacacccag acagccagaa ggcacgcgac
     1441 gagatcaagg aggcctctca gaaggccgag gaggtgaagg agagatgcaa gagcgccagc
     1501 ggaagtggcg gaggaggcga cgtcgctagt gacatcgaga agctgtgcaa gaaggcggaa
     1561 gaggaggcca aggaggccca ggagaaggcc gacgagctga gacaacgaca ccccgattcg
     1621 caggcggcgg aagacgcgga ggatctcgca aatctggctg tcgccgccgt gctgaccgct
     1681 tgtctgctgg cccaggagca tcccaacgcc gatatcgcta agctgtgcat caaggccgcg
     1741 agcgaagctg cggaggccgc gtcaaaagcc gccgagctgg cccaacggca cccagacagt
     1801 caagcagcca gagatgctat caaactcgcc tctcaagccg cccgcgccgt gatcctggcc
     1861 atcatgctcg ccgcagagaa tcccaacgct gatattgcaa agctatgtat taaggcggcc
     1921 agcgaagccg cagaggctgc ctcgaaagct gcggaacttg cacagcggca ccccgactcc
     1981 caggccgccc gagatgcaat taaactggcc agtcaggccg ctgaggcggt ggagagagcc
     2041 atctggctgg ccgccgagaa ccccaatgcc gatatcgcca aaaaatgcat caaagcggcc
     2101 tccgaagcag ccgaggaggc ctctaaagct gccgaagagg ctcaaagaca cccagacagc
     2161 cagaaggcac gcgacgagat caaggaggcc tctcagaagg ccgaggaggt gaaggagaga
     2221 tgcaagagcg ccagcggaag tggcggagga ggcgacgtcg ctagtgacat cgagaagctg
     2281 tgcaagaagg cggaagagga ggccaaggag gcccaggaga aggccgacga gctgagacaa
     2341 cgacaccccg attcgcaggc ggcggaagac gcggaggatc tcgcaaatct ggctgtcgcc
     2401 gccgtgctga ccgcttgtct gctggcccag gagcatccca acgccgatat cgctaagctg
     2461 tgcatcaagg ccgcgagcga agctgcggag gccgcgtcaa aagccgccga gctggcccaa
     2521 cggcacccag acagtcaagc agccagagat gctatcaaac tcgcctctca agccgcccgc
     2581 gccgtgatcc tggccatcat gctcgccgca gagaatccca acgctgatat tgcaaagcta
     2641 tgtattaagg cggccagcga agccgcagag gctgcctcga aagctgcgga acttgcacag
     2701 cggcaccccg actcccaggc cgcccgagat gcaattaaac tggccagtca ggccgctgag
     2761 gcggtggaga gagccatctg gctggccgcc gagaacccca atgccgatat cgccaaaaaa
     2821 tgcatcaaag cggcctccga agcagccgag gaggcctcta aagctgccga agaggctcaa
     2881 agacacccag acagccagaa ggcacgcgac gagatcaagg aggcctctca gaaggccgag
     2941 gaggtgaagg agagatgcaa gagcgccagc ggaagtggcg gaggaggcga cgtcgctagc
     3001 atgctcaaga tggagtctac acagcagatg gctagttcga ttattaacac ttcttttgaa
     3061 gccgcagtcg ttgctgccac ttcaactctt gaacttatgg gcattcaata tgattataat
     3121 gaagtttata ctagagttaa gagcaaattt gattacgtaa tggatgattc aggtgttaaa
     3181 aacaatctgt tgggtaaagc ggcaactatt gatcaggcac tgaatggaaa atttggttca
     3241 gcagcaagaa atagaaattg gatggctgat actagaacta cagctagact agatgaagat
     3301 gtgaacaaac ttagaatgat gctatcatct aaaggaattg atcaaaaaat gagagtactt
     3361 aatgcatgtt tcaatgtgaa gcgagttcca ggtaaatcat catcaataat taaatgtact
     3421 agattaatga gagataagat tgaacgtggt gaagtcgaag tggatgattc gtttgttgaa
     3481 gaaaaaatgg aagttgacac aattgattgg aaatctagat atgaacaatt ggaaaaacga
     3541 tttgaatcgc tgaaacaaag ggtgaatgaa aaatatacaa gttgggtgca aaaagcaaaa
     3601 aaggtaaatg aaaatatgta ctctctccaa aacgttatat cacaacagca gagccagatt
     3661 gctgatttac aaaattactg taataaattg gaagttgatc tacaaaataa aattagttca
     3721 cttgtgtctt cagtcgaatg gtacctgaaa tcaatggaat tgcctgatga aattaagaca
     3781 gatattgaac aacaactaaa ttccattgat gtaatcaatc caattaatgc tatagatgat
     3841 tttgaatcac tgatcagaaa tattatactc gactatgaca gaatattttt aatgtttaaa
     3901 ggattaatga ggcaatgcaa ctatgaatat acgtatgaat aggatccact agtccagtgt
     3961 ggtggaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag ggcccgttta
     4021 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     4081 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     4141 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     4201 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     4261 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     4321 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4381 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4441 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4501 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4561 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4621 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4681 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4741 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4801 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4861 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4921 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4981 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     5041 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5101 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5161 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5221 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5281 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5341 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5401 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5461 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5521 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5581 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5641 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5701 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5761 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5821 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5881 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5941 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     6001 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     6061 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     6121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     6241 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     6301 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     6361 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     6421 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     6481 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     6541 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     6601 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     6661 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6721 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6781 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6841 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6901 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6961 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     7021 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     7081 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     7141 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     7201 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7261 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7321 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7381 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7441 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7501 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7561 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7621 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7681 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7741 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7801 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7861 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7921 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7981 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8041 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8101 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8161 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8221 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8281 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8341 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8401 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtc
//