LOCUS       pVG173                  7983 bp    DNA     circular SYN 06-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7983)
  AUTHORS   Yanfei Zhang
  TITLE     Direct Submission
  JOURNAL   Exported Dec 19, 2025 from SnapGene 8.2.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(12..31)
                     /label=T3_P
     primer_bind     complement(52..68)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(57..178)
                     /label=lac Promoter
                     /note="lac Promoter"
     protein_bind    76..92
                     /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)."
     primer_bind     complement(137..158)
                     /label=pD153_GIB_3_REV
     primer_bind     complement(137..158)
                     /label=Primer 2
     protein_bind    145..166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    complement(1215..2073)
                     /label=AmpR
                     /note="AmpR"
     primer_bind     2052..2071
                     /label=Yfz_Seq25
                     /note="AmpR 5' rev direction seq"
     promoter        complement(2074..2178)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2215..2239
                     /label=Yfz_Oli26
                     /note="R-MluI-Scar out the AscI in plasmid P1"
     primer_bind     complement(2612..2640)
                     /label=Yfz_Oli27
                     /note="F-XhoI-Scar out the SalI in plasmid P1"
     rep_origin      complement(2815..3270)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3411..3427
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3434..3452
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    complement(3491..4026)
                     /label=ScHIS3 downstream flanking (+1/+536)
     misc_feature    complement(3491..3809)
                     /label=ScHIS3 3' end
                     /note="ScHIS3 3' end"
     primer_bind     4007..4037
                     /label=YfzOli29
                     /note="SacI_insert Fragment_PstI-R Can be used 
                     forsequencing"
     primer_bind     4039..4050
                     /label=Forward EL222 primer
     misc_feature    complement(4101..4115)
                     /label=SGI-S1
                     /note="SGI-S1"
     misc_feature    4114..4827
                     /label=PGK1pr VG
                     /note="Annotated using Yeastgenome database"
     misc_feature    4116..4833
                     /label=PGK1 Promoter
                     /note="PGK1 Promoter"
     primer_bind     4824..4837
                     /label=Forward EL222 primer
     misc_feature    4844..4856
                     /label=KOZAK
                     /note="KOZAK"
     CDS             4860..4880
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    4881..5114
                     /label=VP16 Binding Domain
     primer_bind     4949..4965
                     /label=VP16 fwd
     misc_feature    5121..5750
                     /label=EL122
     primer_bind     complement(5380..5403)
                     /label=middle EL222 reverse primer
     misc_feature    5751..5756
                     /label=pJLA Homology Arm
     misc_feature    complement(5751..5756)
                     /label=MCS
                     /note="poly-linker region"
     misc_feature    5763..5768
                     /label=STOP
                     /note="STOP"
     primer_bind     complement(5767..5790)
                     /label=Reverse EL222
     misc_feature    5769..6020
                     /label=CYC1 Terminator
                     /note="CYC1 Terminator"
     primer_bind     complement(5787..5803)
                     /label=CYC1term rev
     misc_feature    6021..6050
                     /label=SGI-S2
                     /note="SGI-S2"
     primer_bind     complement(6025..6061)
                     /label=pD153_GIB_3_REV
     primer_bind     complement(6043..6067)
                     /label=Primer 2
     misc_feature    6051..6063
                     /label=SGI-S2
                     /note="SGI-S2"
     misc_feature    6069..7482
                     /label=CG HIS3 w promoter and terminator
     misc_feature    complement(6069..6735)
                     /label=Cg HIS3 terminator
     primer_bind     complement(6204..6223)
                     /label=Yfz_Seq24
                     /note="IntSeq_CgHIS3 terminator_middle_forward"
     misc_feature    complement(6736..7368)
                     /label=CgHIS3
                     /note="Candida glabrata imidazoleglycerol-phosphate
                     dehydratase (HIS3) gene, complete cds"
     gene            6800..7429
                     /label=CgHIS3
                     /note="CgHIS3"
     primer_bind     complement(7106..7125)
                     /label=Yfz_Seq23
                     /note="IntSeq_CgHIS3_middle_forward"
     misc_feature    complement(7369..7482)
                     /label=CgHIS3 promoter
     misc_feature    7446..7946
                     /label=ScHIS3 5' end
                     /note="ScHIS3 5' end"
     misc_feature    complement(7483..7946)
                     /label=ScHIS upstream flanking (-464/-1)
     promoter        complement(7483..7670)
                     /gene="S. cerevisiae HIS3"
                     /label=ScHIS3 promoter
BASE COUNT         2152 a         1841 c         1830 g         2160 t
ORIGIN
        1 cagcttttgt tccctttagt gagggttaat tccgagcttg gcgtaatcat ggtcatagct
       61 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacataggag ccggaagcat
      121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      241 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
      301 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      361 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      421 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
      481 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      541 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      601 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      661 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      721 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      781 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      841 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      901 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      961 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1021 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1081 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1141 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1201 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1261 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     1321 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     1381 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     1441 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     1501 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     1561 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     1621 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     1681 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     1741 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     1801 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     1861 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     1921 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     1981 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2041 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2101 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2161 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcc tcggcgcgtg
     2221 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     2281 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     2341 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     2401 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     2461 tgagagtgca ccataccacc ttttcaattc atcatttttt ttttattctt ttttttgatt
     2521 tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
     2581 ggagcacaga cttagattgg tatatatacg catattgggt cagcagtaca gaaccctcga
     2641 cgaagttgta agtggtggat cagatatcat cattgttggc agaggacttt tcgccaaggg
     2701 tagagatcct aatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     2761 cctatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     2821 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     2881 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     2941 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3001 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3061 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3121 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3181 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3241 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     3301 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     3361 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     3421 ggccagtgaa ttgtaatacg actcactata gggggccagc gtggccgttt aaacatttaa
     3481 atggccggcc gctgccaccg ttgtttccac cgaagaaacc accgttgccg taaccaccac
     3541 gacggttgtt gctaaagaag ctgccaccgc cacggccacc gttgtagccg ccgttgttgt
     3601 tattgtagtt gctactgtta tttctggcac ttcttggttt tcctcttaag tgaggaggaa
     3661 cataaccatt ctcgttgttg tcgttgatgc ttaaattttg cacttgttcg ctcagttcag
     3721 ccataatatg aaatgctttt cttgttgttc ttacggaata ccacttgcca cctatcacca
     3781 caactaactt tttcccgttc ctccatctct tttatatttt ttttctggag ttcaagagaa
     3841 aaaaaaagaa aaagcaaaaa gaaaaaagga aagcgcgcct cgttcagaat gacacgtata
     3901 gaatgatgca ttaccttgtc atcttcagta tcatactgtt cgtatacata cttactgaca
     3961 ttcataggta tacatatata cacatgtata tatatcgtat gctgcagctt taaataatcg
     4021 gtgtcaagat ctgagctcca ccgcggtggc ggccgcttat ttactcagga tcccgcatgt
     4081 tgcaggtggg agttccagct cccgggaaac agctgcaaga attactcgtg agtaaggaaa
     4141 gagtgaggaa ctatcgcata cctgcattta aagatgccga tttgggcgcg aatcctttat
     4201 tttggcttca ccctcatact attatcaggg ccagaaaaag gaagtgtttc cctccttctt
     4261 gaattgatgt taccctcata aagcacgtgg cctcttatcg agaaagaaat taccgtcgct
     4321 cgtgatttgt ttgcaaaaag aacaaaactg aaaaaaccca gacacgctcg acttcctgtc
     4381 ttcctattga ttgcagcttc caatttcgtc acacaacaag gtcctagcga cggctcacag
     4441 gttttgtaac aagcaatcga aggttctgga atggcgggaa agggtttagt accacatgct
     4501 atgatgccca ctgtgatctc cagagcaaag ttcgttcgat cgtactgtta ctctctctct
     4561 ttcaaacaga attgtccgaa tcgtgtgaca acaacagcct gttctcacac actcttttct
     4621 tctaaccaag ggggtggttt agtttagtag aacctcgtga aacttacatt tacatatata
     4681 taaacttgca taaattggtc aatgcaagaa atacatattt ggtcttttct aattcgtagt
     4741 ttttcaagtt cttagatgct ttctttttct cttttttaca gatcatcaag gaagtaatta
     4801 tctacttttt acaacaaata taaaacaccg cggtggcggc cgctaaaatc atggctagcc
     4861 cgaaaaaaaa gagaaaagta gccccaccaa cggacgtttc acttggtgac gaattacatt
     4921 tagacggtga ggatgtagct atggcacatg ccgatgctct tgatgatttt gatctggaca
     4981 tgttaggtga tggggattct cccggccccg gatttacacc acatgattct gctccgtatg
     5041 gcgcgttgga catggcagat ttcgaatttg aacagatgtt tactgatgca cttggtatcg
     5101 acgaatatgg aggtgaattc ggagcagatg atactagagt tgaagtccaa cctccagccc
     5161 aatgggtatt ggatttgatc gaggcctcac ccatagcaag tgttgttagt gatcccagat
     5221 tggctgataa tcctttgatt gctatcaacc aggcctttac cgatctaact gggtattctg
     5281 aggaagaatg tgttggtcgt aattgtagat ttttggcaag atctggtact gaaccctggc
     5341 taacagataa gatccgtcaa ggtgttcgtg agcataaacc cgtgttggtt gagattttga
     5401 attacaaaaa ggatggtact ccttttcgta atgcagtttt ggtggcacca atctatgatg
     5461 acgatgatga attactatac ttccttggta gccaagtgga agtggatgac gatcaaccga
     5521 acatgggcat ggcccgtaga gaacgtgctg cggagatgtt aaaaactctt tcaccacgtc
     5581 aattagaagt cacaactctg gttgcatcag gcctaagaaa taaagaagtg gctgccagac
     5641 tgggtctttc agaaaaaact gtcaaaatgc acagaggttt ggtaatggaa aaattaaact
     5701 taaaaacgag tgcagattta gttagaattg ccgtagaagc aggtatttaa gtcgacctcg
     5761 agtaatgatc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     5821 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     5881 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     5941 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     6001 gaaggcttta atttgcggcc ttatcgccgg cgaataaatt aaggcgcgcc aacatatccg
     6061 gaggtaccca aagccgaatc caccacggtc accgttgttt ctgaagaaac caccacggcc
     6121 gccaccgttg ttgaaaccac cacgaggagc agaatcgcct cttctgccac caccgttacg
     6181 gatgtgaggt ggaacgtaac caccacgtct ttgttgccct tcaggcatgg aaacttgctc
     6241 tgctaactca gtcatattgg atatatgtat tactttcttt attttttgta gtttagtctt
     6301 ctatttcttt tatagtttgg tatctttttg caaaaaattt acctctatat cgtgttcgtg
     6361 cgtatttctt cctctataat atgaaaagaa aggcagtagt cttggttttc tattattcca
     6421 aatattacaa gaatacccag aaccacaaga agcagaagaa actaactgct ttatatactt
     6481 aatttcccgc cacaagctca tcgcccagaa gtttttttca gtttttcacc tgacaatctg
     6541 gcagctcgct gaaaaatttt cacctggggt cgcctgctac gtagagtgat cacgtttatg
     6601 aatagcacat tatatagtgg cccagacggt ctgatggtag ttcatgtagt taagaataaa
     6661 tactctgttt gtgtgatttc tatgtgtacg ttatctatat aaatattgtt gatggtggta
     6721 gctgtgggct gtgttctatg ctaggacacc cttagtggat ggaacgtcat cgcgacccgt
     6781 aggtgttctt gcctctctga tggcgatggc gagcgccttg aaagctgatt cggagcggtg
     6841 gtggtcgttg gtgccccgca gacagtccac atgcatggtg atacgcgcct cagtggcgaa
     6901 actctccaag aagtgcggga tcatctctgt ggatagctgg cctatgcgtt ctcgcttcag
     6961 gcccagctcc accacggcga aaggcctatt ggagaagtca accacagcgc ggctcagcgc
     7021 ctcgtccagt ggtgcaaacc catgcccgaa cctcttgata ccacgcacgg agccaagcgc
     7081 ctccttgaac gcttggccca gcgcaatccc acagtcctca acagtgtggt ggtcatcgat
     7141 gtgcaagtcc cctatacact ccaggatcag agaccacccg ccatgcttcg ccagcgcagt
     7201 caacatgtgg tccaaaaacc caactccagt gtgcacatgg atggtctgcc catcaccact
     7261 agcatattcc ttgcccagta tgctctcccg tacagaaaca gacccaccgt caagatccag
     7321 cgccagctgt atattcgtct cctgcgtaac cctcttaaca aacgccatcc tttgctcgat
     7381 gcttctcttt gcttcaatta tctcaacttt aagtaaacct gaagtaaact agaactagat
     7441 taaatcagaa ggcatccgca gctttatata catatagagc ctctttgcct tcgtttatct
     7501 tgcctgctca ttttttagta tattcttcga agaaatcaca ttactttata taatgtataa
     7561 ttcattatgt gataatgcca atcgctaaga aaaaaaaaga gtcatccgct aggtggaaaa
     7621 aaaaaaatga aaatcattac cgaggcataa aaaaatatag agtgtactag aggaggccaa
     7681 gagtaataga aaaagaaaat tgcgggaaag gactgtgtta tgacttccct gactaatgcc
     7741 gtgttcaaac gatacctggc agtgactcct agcgctcacc aagctcttaa aacgagaatt
     7801 aagaaaaagt cgtcatcttt cgataagttt ttcccacagc aaagcaatag tagaaaaaaa
     7861 caatgggaaa cgttgaatga agacaaagcg tcgtggttta aaaggaaata cgctcacgta
     7921 catgctaggg aacaggaccg tgcagcggcc agcgtggccg tttaaacatt taaatggccg
     7981 gcc
//
