LOCUS       Exported                8604 bp ds-DNA     circular SYN 26-JUN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8604)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jun 6, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     f1:gGAAAATCTCTACTTCCAGATGggcgcgcctatgaaagcgttaacggccaggcaacaagag
            r1:CCCCCTCGAGGTCGACGGTATCGATAAGCTTTAAACGCGTttagAGCTTCCCACCGTAC
FEATURES             Location/Qualifiers
     source          1..8604
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    313..508
                     /locus_tag="HIS promoter"
                     /label=HIS promoter
     misc_feature    509..1168
                     /locus_tag="HIS3"
                     /label=HIS3
     misc_feature    1429..1525
                     /locus_tag="f1 ori"
                     /label=f1 ori
     misc_feature    1523..1525
                     /locus_tag="PsiI"
                     /label=PsiI
     misc_feature    1526..1533
                     /locus_tag="NotI"
                     /label=NotI
     misc_feature    1778..1783
                     /locus_tag="XbaI"
                     /label=XbaI
     misc_feature    1797..2639
                     /locus_tag="YFP"
                     /label=YFP
     misc_feature    2640..2729
                     /locus_tag="2X Protease resistant linker"
                     /label=2X Protease resistant linker
     misc_feature    2730..3441
                     /locus_tag="URA3"
                     /label=URA3
     misc_feature    3528..3533
                     /locus_tag="STOP"
                     /label=STOP
     misc_feature    3559..3808
                     /locus_tag="CYC1 terminator"
                     /label=CYC1 terminator
     misc_feature    3809..3811
                     /locus_tag="PsiI(1)"
                     /label=PsiI
     misc_feature    3809..3811
                     /locus_tag="f1 ori(2)"
                     /label=f1 ori
     misc_feature    3812..4167
                     /locus_tag="f1 ori(1)"
                     /label=f1 ori
     misc_feature    4183..4331
                     /locus_tag="lacZ_a"
                     /label=lacZ_a
     misc_feature    4398..4460
                     /locus_tag="New Feature"
                     /label=New Feature
     misc_feature    4461..4962
                     /locus_tag="ACT1"
                     /label=ACT1
     misc_feature    4986..4991
                     /locus_tag="EcoRI"
                     /label=EcoRI
     misc_feature    4992..4997
                     /locus_tag="KpnI"
                     /label=KpnI
     misc_feature    4998..5000
                     /locus_tag="START codon"
                     /label=START codon
     misc_feature    5001..5606
                     /locus_tag="LexA-ER-haVP16(1)"
                     /label=LexA-ER-haVP16
     misc_feature    5001..5606
                     /locus_tag="LexA"
                     /label=LexA
     misc_feature    5607..6028
                     /locus_tag="LexA-ER-haVP16"
                     /label=LexA-ER-haVP16
     misc_feature    5607..6028
                     /locus_tag="VP16"
                     /label=VP16
     misc_feature    6030..6035
                     /locus_tag="NheI"
                     /label=NheI
     misc_feature    6036..6041
                     /locus_tag="HpaI"
                     /label=HpaI
     misc_feature    6042..6047
                     /locus_tag="AgeI"
                     /label=AgeI
     misc_feature    6048..6212
                     /locus_tag="cytoplasmic"
                     /label=cytoplasmic
     misc_feature    6156..6161
                     /locus_tag="TeNT cleavage"
                     /label=TeNT cleavage
     misc_feature    6213..6278
                     /label=Transmembrane domain
     primer_bind     6213..6238
                     /label=frag_2_fwd
     misc_feature    6279..6281
                     /locus_tag="stop"
                     /label=stop
     misc_feature    6282..6287
                     /locus_tag="HindIII"
                     /label=HindIII
     misc_feature    6288..6293
                     /locus_tag="ClaI "
                     /label=ClaI
     misc_feature    6303..6308
                     /locus_tag="XhoI "
                     /label=XhoI
     primer_bind     complement(6317..6333)
                     /label=frag_2_Rev
     misc_feature    6780..7399
                     /locus_tag="pBR322 origin"
                     /label=pBR322 origin
     misc_feature    7554..8414
                     /locus_tag="Ampicillin R"
                     /label=Ampicillin R
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg ttttaagagc ttggtgagcg ctaggagtca ctgccaggta tcgtttgaac
      241 acggcattag tcagggaagt cataacacag tcctttcccg caattttctt tttctattac
      301 tcttggcctc ctctagtaca ctctatattt ttttatgcct cttaattaag gtaatgattt
      361 tcattttttt ttttccacct agcggatgac tctttttttt tcttagcgat tggcattatc
      421 acataatgaa ttatacatta tataaagtaa tgtgatttct tcgaagaata tactaaaaaa
      481 tgagcaggca agataaacga aggcaaagat gacagagcag aaagccctag taaagcgtat
      541 tacaaatgaa accaagattc agattgcgat ctctttaaag ggtggtcccc tagcgataga
      601 gcactcgatc ttcccagaaa aagaggcaga agcagtagca gaacaggcca cacaatcgca
      661 agtgattaac gtccacacag gtatagggtt tctggaccac atgatacatg ctctggccaa
      721 gcattccggc tggtcgctaa tcgttgagtg cattggtgac ttacacatag acgaccatca
      781 caccactgaa gactgcggga ttgctctcgg tcaggctttt aaagaggccc tactggcgcg
      841 tggagtaaaa aggtttggat caggatttgc gcctttggat gaggcacttt ccagagcggt
      901 ggtagatctt tcgaacaggc cgtacgcagt tgtcgaactt ggtttgcaaa gggagaaagt
      961 aggagatctc tcttgcgaga tgatcccgca ttttcttgaa agttttgcag aggctagcag
     1021 aattaccctc cacgttgatt gtctgcgagg caagaatgat catcaccgta gtgagagtgc
     1081 gttcaaggct cttgcggttg ccataagaga agccacctcg cccaatggaa ccaacgatgt
     1141 tccctccacc aaaggtgttc ttatgtagtg acaccgatta tttaaagctg cagcatacga
     1201 tatatataca tgtgtatata tgtataccta tgaatgtcag taagtatgta tacgaacagt
     1261 atgatactga agatgacaag gtaatgcatc attctatacg tgtcattctg aacgaggcgc
     1321 gctttccttt tttctttttg ctttttcttt ttttttctct tgaactcgac ggatcatatg
     1381 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaaacg
     1441 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1501 aggccgaaat cggcaaaatc ccttagcggc cgcaataata tataaacctg tataatataa
     1561 ccttgaagac tatatttctt ttcgagctcc ctaggtgctg tatatactca cagcataact
     1621 gtatatacac ccagggtcta ggtgctgtat atactcacag cataactgta tatacaccca
     1681 gggtctaggt gctgtatata ctcacagcat aactgtatat acacccaggg tctaggtgct
     1741 gtatatactc acagcataac tgtatataca cccagggtct agagcatgtg ctctgtatgt
     1801 atataaaact cttgttttct tcttttctct tctttcctta tacattagga cctttgcagc
     1861 ataaattact atacttctat actagtggat cccccgggct gcaggaattc gatatcaagc
     1921 ttatgagatc taaaggtgaa gaattattca ctggtgttgt cccaattttg gttgaattag
     1981 atggtgatgt taatggtcac aaattttctg tctccggtga aggtgaaggt gatgctactt
     2041 acggtaaatt gaccttaaaa tttatttgta ctactggtaa attgccagtt ccatggccaa
     2101 ccttagtcac tactttaggt tatggtttga tgtgttttgc tagataccca gatcatatga
     2161 aacaacatga ctttttcaag tctgccatgc cagaaggtta tgttcaagaa agaactattt
     2221 ttttcaaaga tgacggtaac tacaagacca gagctgaagt caagtttgaa ggtgatacct
     2281 tagttaatag aatcgaatta aaaggtattg attttaaaga agatggtaac attttaggtc
     2341 acaaattgga atacaactat aactctcaca atgtttacat catggctgac aaacaaaaga
     2401 atggtatcaa agttaacttc aaaattagac acaacattga agatggttct gttcaattag
     2461 ctgaccatta tcaacaaaat actccaattg gtgatggtcc agtcttgtta ccagacaacc
     2521 attacttatc ctatcaatct aagttatcca aagatccaaa cgaaaagaga gaccacatgg
     2581 tcttgttaga atttgttact gctgctggta ttacccatgg tatggatgaa ttgtacaaag
     2641 gttccacgtc aggttctggc aagcccggat caggcgaggg tagcggctct acttctgggt
     2701 ctggtaagcc tgggagtgga gaggggagct cgaaagctac atataaggaa cgtgctgcta
     2761 ctcatcctag tcctgttgct gccaagctat ttaatatcat gcacgaaaag caaacaaact
     2821 tgtgtgcttc attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag
     2881 gtcccaaaat ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg
     2941 gcacagttaa gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca
     3001 gaaaatttgc tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa
     3061 tagcagaatg ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg
     3121 gtttgaagca ggcggcggaa gaagtaacaa aggaacctag aggtcttttg atgttagcag
     3181 aattgtcatg caagggctcc ctagctactg gagaatatac taagggtact gttgacattg
     3241 cgaagagcga caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag
     3301 atgaaggtta cgattggttg attatgacac ccggtgtggg tttagatgac aagggagacg
     3361 cattgggtca acagtataga accgtggatg atgtggtctc tacaggatct gacattatta
     3421 ttgttggaag aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca
     3481 gaaaagcagg ctgggaagca tatttgagaa gatgcggcca gcaaaactaa tagacaggcc
     3541 ccttttcctt tgtctatatc atgtaattag ttatgtcacg cttacattca cgccctcctc
     3601 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
     3661 atttttttta atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt
     3721 ttttctgtac aaacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt
     3781 tttgggacgc tcgaaggctt taatttgcta aatcaaaaga atagaccgag atagggttga
     3841 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3901 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3961 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     4021 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     4081 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     4141 cgcttaatgc gccgctacag ggcgcgtcgc gccattcgcc attcaggctg cgcaactgtt
     4201 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaag gggggatgtg
     4261 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     4321 cggccagtga attgtaatac gactcactat agggcgaatt ggagaaacag cggccgccta
     4381 tagggcgaat tggagctcca ccgcggtggc ggccgctcta gagaagcggg taagctgcca
     4441 cagcaattaa tgcacaacat ttaacctaca ttcttcctta tcggatcctc aaaaccctta
     4501 aaaacatatg cctcacccta acatattttc caattaaccc tcaatatttc tctgtcaccc
     4561 ggcctctatt ttccattttc ttctttaccc gccacgcgtt tttttctttc aaattttttt
     4621 cttccttctt ctttttcttc cacgtcctct tgcataaata aataaaccgt tttgaaacca
     4681 aactcgcctc tctctctcct ttttgaaata tttttgggtt tgtttgatcc tttccttccc
     4741 aatctctctt gtttaatata tattcattta tatcacgctc tctttttatc ttcctttttt
     4801 tcctctctct tgtattcttc cttccccttt ctactcaaac caagaagaaa aagaaaaggt
     4861 caatctttgt taaagaatag gatcttctac tacatcagct tttagatttt tcacgcttac
     4921 tgcttttttc ttcccaagat cgaaaattta ctgaattaac aactagtgga tcccccgggc
     4981 tgcaggaatt cggtaccatg aaagcgttaa cggccaggca acaagaggtg tttgatctca
     5041 tccgtgatca catcagccag acaggtatgc cgccgacgcg tgcggaaatc gcgcagcgtt
     5101 tggggttccg ttccccaaac gcggctgaag aacatctgaa ggcgctggca cgcaaaggcg
     5161 ttattgaaat tgtttccggc gcatcacgcg ggattcgtct gttgcaggaa gaggaagaag
     5221 ggttgccgct ggtaggtcgt gtggctgccg gtgaaccact tctggcgcaa cagcatattg
     5281 aaggtcatta tcaggtcgat ccttccttat tcaagccgaa tgctgatttc ctgctgcgcg
     5341 tcagcgggat gtcgatgaaa gacatcggca ttatggatgg tgacttgctg gcagtgcata
     5401 aaactcagga tgtacgtaac ggtcaggtcg ttgtcgcacg tattgatgac gaggttaccg
     5461 ttaagcgcct gaaaaaacag ggcaataaag tcgaactgtt gccagaaaat agcgagttta
     5521 aaccaattgt cgtagatctt cgtcagcaga gcttcaccat tgaagggctg gcggttgggg
     5581 ttattcgcaa cggcgactgg ctgggtgata tcatgcagga tctgccgggc aacgataaca
     5641 gcaccgcggg cgcgcgtacg aaaaacaatt acgggtctac catcgagggc ctgctcgatc
     5701 tcccggacga cgacgccccc gaagaggcgg ggctggcggc tccgcgcctg tcctttctcc
     5761 ccgcgggaca cacgcgcaga ctgtcgacgg cccccccgac cgatgtcagc ctgggggacg
     5821 aattgcactt agacggcgag gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg
     5881 atctggacat gttgggggac ggggattccc cgggtccggg atttaccccc cacgactccg
     5941 ccccctacgg cgctctggat atggccgact tcgagtttga gcagatgttt accgatgccc
     6001 ttggaattga cgagtacggt gggaagctcg ctagcgttaa caccggtgat gaggtggtgg
     6061 acatcatgag ggtgaatgtg gacaaggtcc tggagcggga ccagaagttg tcggagctgg
     6121 atgaccgtgc agatgccctc caggcagggg cctcccagtt tgaaacaagt gcagccaagc
     6181 tcaagcgcaa atactggtgg aaaaacctca agatgatgat catcttggga gtgatctgcg
     6241 ccatcatcct catcatcatc atcgtttact tcagcactta aaagcttatc gataccgtcg
     6301 acctcgaggg ggggcccggt acccagcttc ggccgtgtac ccagcttttg ttccctttag
     6361 tgagggttaa ttccgagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6421 tatccgctca caattccaca caacatagga gccggaagca taaagtgtaa agcctggggt
     6481 gcctaatgag tgaggtaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6541 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6601 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6661 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6721 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6781 gcgttgctgg cgtttttcca taggctcggc ccccctgacg agcatcacaa aaatcgacgc
     6841 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt cccccctgga
     6901 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6961 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     7021 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     7081 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     7141 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     7201 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     7261 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     7321 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     7381 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     7441 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7501 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7561 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7621 tgactgcccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7681 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7741 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7801 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7861 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7921 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgaaaaaa agcggttagc
     7981 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8041 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8101 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8161 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8221 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8281 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8341 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8401 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8461 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     8521 acatttcccc gaaaagtgcc acctgacgtc ttattatcat gacattaacc tataaaaata
     8581 ggcgtatcac gaggcccttt cgtc
//
