LOCUS       Exported                5283 bp ds-DNA     circular SYN 11-JAN-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pROS13-Phleo-TPS1_GSY2-clone5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5283)
  AUTHORS   Haoqi Chen
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 11, 2023 from SnapGene Viewer 3.1.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5283
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(522..1110)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1281..2141)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2142..2246)
                     /gene="bla"
                     /note="AmpR promoter"
     terminator      2393..2590
                     /note="TEF terminator"
                     /note="Ashbya gossypii TEF terminator"
     misc_feature    2596..2976
                     /note="ble (Phleo)"
     promoter        complement(2977..3320)
                     /note="TEF promoter"
                     /note="Ashbya gossypii TEF promoter"
     promoter        3475..3743
                     /note="SNR52 promoter"
                     /note="promoter for the S. cerevisiae small nucleolar RNA 
                     gene SNR52"
     misc_feature    3744..3763
                     /note="GSY2_seed"
     misc_RNA        3764..3838
                     /note="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      3843..3862
                     /note="SUP4 terminator"
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     rep_origin      4087..5068
                     /note="2u ori"
                     /note="yeast 2u plasmid origin of replication"
     terminator      5114..5133
                     /note="SUP4 terminator"
                     /note="transcription terminator for the S. cerevisiae SUP4 
                     tRNA gene"
     misc_RNA        complement(5138..5212)
                     /note="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    5213..5232
                     /note="TPS1_seed"
     misc_feature    complement(5233..218)
                     /note="SNR52 promoter"
ORIGIN
        1 caactttcac ttctacagcg tttgaccaaa atcttttgaa cagaacattg tagggtgtga
       61 aaaaatgcgc acctttaccg ctagcccaag agggcactac aaaatctaga gttgtacttc
      121 aaacgtacat gtaatcacct tgtatatact cgaaagaaaa catcaagttt ctgtataaat
      181 atgagtgaaa gcataatcat acattatctt ttcaaagaag ctccagcttt tgttcccttt
      241 agtgagggta ttcacgtaga cggataggta tagccagaca tcagcagcat acttcgggaa
      301 ccgtaggctc tagagagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
      361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
      421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
      481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
      541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
      601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
      661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
      721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
      781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
      841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
      901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
      961 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     1141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     1201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     1261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     1321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     1381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     1441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     1501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     1561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     1621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     1681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     1741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     1801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     1861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     1921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     2101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     2161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2221 aaaaataaac aaataggggt tccgcgcact cgagactata tgtgaaggca tggctatggc
     2281 acggcagaca ttccgccaga tcatcaatag gcaccttcgt acgctgcagg attaagggtt
     2341 ctcgagagct cgttttcgac actggatggc ggcgttagta tcgaatcgac agcagtatag
     2401 cgaccagcat tcacatacga ttgacgcatg atattacttt ctgcgcactt aacttcgcat
     2461 ctgggcagat gatgtcgagg cgaaaaaaaa tataaatcac gctaacattt gattaaaata
     2521 gaacaactac aatataaaaa aactatacaa atgacaagtt cttgaaaaca agaatctttt
     2581 tattgtcagt actgatcatg agatgcctgc aagcaattcg ttctgtatca ggcgcaggag
     2641 cgtcccgtcc gggtcgacca aagcggccat cgtgcctccc cactcctgca gttcgggggc
     2701 atggatgcgc ggatagccgc tgctggtttc ctggatgccg acggatttgc actgccggta
     2761 gaactccgcg aggtcgtcca gcctcaggca gcagctgaac caactcgcga ggggatcgag
     2821 cccggggtgg gcgaagaact ccagcttgag atccccgcgc tggaggatca tccagccggc
     2881 gtcccggaaa acgattccga agcccaacct ttcatagaag gcggccgtgg aatcgaaatc
     2941 tcgtgatggc aggttgggcg tcgcttggtc ggccatggtt gtttatgttc ggatgtgatg
     3001 tgagaactgt atcctagcaa gattttaaaa ggaagtatat gaaagaagaa cctcagtggc
     3061 aaatcctaac cttttatatt tctctacagg ggcgcggcgt ggggacaatt caacgcgtct
     3121 gtgaggggag cgtttccctg ctcgcaggtc tgcagcgagg agccgtaatt tttgcttcgc
     3181 gccgtgcggc catcaaaatg tatggatgca aatgattata catggggatg tatgggctaa
     3241 atgtacgggc gacagtcaca tcatgcccct gagctgcgca cgtcaagact gtcaaggagg
     3301 gtattctggg cctccatgtc gctggccggg tgacccggcg gggacgaggc aagctaaaca
     3361 gatctctaga cctatggcct atgccacctt tcgagaggac gatgcccgtg tctaaatgat
     3421 tcgaccagcc taagaatgtt caacccctca ctaaagggaa caaaagctgg agcttctttg
     3481 aaaagataat gtatgattat gctttcactc atatttatac agaaacttga tgttttcttt
     3541 cgagtatata caaggtgatt acatgtacgt ttgaagtaca actctagatt ttgtagtgcc
     3601 ctcttgggct agcggtaaag gtgcgcattt tttcacaccc tacaatgttc tgttcaaaag
     3661 attttggtca aacgctgtag aagtgaaagt tggtgcgcat gtttcggcgt tcgaaacttc
     3721 tccgcagtga aagataaatg atcaatttgt aaaaaagaca agagttttag agctagaaat
     3781 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtggt
     3841 gctttttttg ttttttatgt cttcgagtca tgtaattagt tatgtcacgc ttacgttcac
     3901 gccctccacg catttaagca taaacacgca ctatgccgtt cttctcatgt atatatatat
     3961 acaggcaaca cgcagatata ggtgcgacgt gaacagtgag ctgtatgtgc gcagctcgcg
     4021 ttgcattttc ggaagcgctc gttttcggaa acgctttgaa gttcctattc cgaagttcct
     4081 attctgtaga aagtatagga acctcagagc gcttttgaaa accaaaagcg ctctgaagac
     4141 gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata
     4201 ccgcttccac aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc
     4261 tatataacct acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca
     4321 ttttttatgt ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc
     4381 atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga
     4441 caaaatagaa gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact
     4501 ttctgttcac aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta
     4561 tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag
     4621 gcttttttta tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc
     4681 tacagtgcaa aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta
     4741 atctaagatg ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa
     4801 gaagtataga ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa
     4861 atgcagctca gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca
     4921 aaaatgaagc acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg
     4981 taaaaatgca gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt
     5041 ctacaaaatg aagcacagat gcttcgttgg agggcgtgaa cgtaagcgtg acataactaa
     5101 ttacatgact cgaagacata aaaaacaaaa aaagcaccac cgactcggtg ccactttttc
     5161 aagttgataa cggactagcc ttattttaac ttgctatttc tagctctaaa actgaatggt
     5221 gctattattg tagatcattt atctttcact gcggagaagt ttcgaacgcc gaaacatgcg
     5281 cac
//
