LOCUS       Exported                5632 bp ds-DNA     circular SYN 26-NOV-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSC101-GFPmut2-mScarlet-I TGA
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5632)
  AUTHORS   Mariliis Hinnu
  TITLE     Direct Submission
  JOURNAL   Exported Monday, August 28, 2023 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5632
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             complement(196..1056)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1057..1161)
                     /gene="bla"
                     /note="AmpR promoter"
     misc_feature    1213..1240
                     /note="rrnB T2 terminator"
     misc_feature    1361..1366
                     /note="-35"
     misc_feature    1380..1385
                     /note="-10"
     misc_feature    1395
                     /note="dnaKp1"
                     /note="transcription start of promoter P1 (rpoH dependent)"
     misc_feature    1446..1511
                     /note="Tet_prom_to_SD"
     promoter        1448..1476
                     /note="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             1540..2265
                     /codon_start=1
                     /product="GFP"
                     /note="GFPmut2"
                     /translation="MKGSSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYIMADKQKDG
                     IKVNFKIRHNIEDGSVQLADHYQRNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITHGMDELYK"
     CDS             1552..1559
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /note="yeGFP"
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SK"
     CDS             2283..2984
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /note="mScarlet-I"
                     /translation="MVSKG*SEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQ
                     TAKLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFED
                     GGAVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGD
                     IKMALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEG
                     RHSTGGMDELYK"
     CDS             3087..3110
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /note="8xHis"
                     /translation="HHHHHHHH"
     misc_feature    3144..3145
                     /note="trnscr_start"
                     /note="Tac promoter transcription start site"
     terminator      3195..3289
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     rep_origin      3781..4003
                     /note="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4051..5001
                     /codon_start=1
                     /gene="rep101"
                     /product="protein needed for replication with the pSC101 
                     origin"
                     /note="Rep101"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(5536..5622)
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 agcttgcatg cccaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
       61 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
      121 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
      181 gcgtccggcg tagagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
      241 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
      301 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
      361 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
      421 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
      481 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
      541 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
      601 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
      661 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
      721 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
      781 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
      841 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
      901 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
      961 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     1021 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     1081 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1141 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1201 cattattatc ataaaaggcc atccgtcagg atggccttct atgaatgcct tggctgcgat
     1261 tcattcttta tatgaataaa attgctgtca attttacgtc ttgtcctgcc atatcgcgaa
     1321 atttctgcgc aaaagcacaa aaaatttttg catctccccc ttgatgacgt ggtttacgac
     1381 cccatttagt agtcaaccgc agtgagtgag tctgacgagg ccctttcgtc ttcaagaatt
     1441 ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt
     1501 gctaacgcag tccggggaat tcaggcttgg aggaacaata tgaaaggatc ctctaaaggt
     1561 gaagaattat tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     1621 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     1681 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc
     1741 gcgtatggtc ttcaatgctt tgcgagatac ccagatcata tgaaacagca tgactttttc
     1801 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     1861 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     1921 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac
     1981 tataactcac acaaagtata catcatggca gacaaacaaa aggatggaat caaagttaac
     2041 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacga
     2101 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     2161 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     2221 acagctgctg ggattacaca tggtatggat gaattgtaca aataaccgcc aggaggaaca
     2281 atatggtgag caagggctga agtgaggcag tgatcaagga gttcatgcgg ttcaaggtgc
     2341 acatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     2401 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttct
     2461 cctgggacat cctgtcccct cagttcatgt acggctccag ggccttcatc aagcaccccg
     2521 ccgacatccc cgactactat aagcagtcct tccccgaggg cttcaagtgg gagcgcgtga
     2581 tgaacttcga ggacggcggc gccgtgaccg tgacccagga cacctccctg gaggacggca
     2641 ccctgatcta caaggtgaag ctccgcggca ccaacttccc tcctgacggc cccgtaatgc
     2701 agaagaagac aatgggctgg gaagcgtcca ccgagcggtt gtaccccgag gacggcgtgc
     2761 tgaagggcga cattaagatg gccctgcgcc tgaaggacgg cggccgctac ctggcggact
     2821 tcaagaccac ctacaaggcc aagaagcccg tgcagatgcc cggcgcctac aacgtcgacc
     2881 gcaagttgga catcacctcc cacaacgagg actacaccgt ggtggaacag tacgaacgct
     2941 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtgaccg cctaatgagc
     3001 gggctttaat cactagtgaa ttcgcggccg cctgcaggtc gaccatatgg gagagctccg
     3061 tcgacaagct tgcggccgca ctcgagcacc accaccacca ccaccaccac taattgatta
     3121 atacctaggc tgctaaacaa agcccatgga tcgatagctg gtcgacagga ctctggggtt
     3181 cgagagctcg cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga
     3241 tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc caagcactag
     3301 ggacagtaag acgggtaagc ctgttgatga taccgctgcc ttactgggtg cattagccag
     3361 tctgaatgac ctgtcacggg ataatccgaa gtggtcagac tggaaaatca gagggcagga
     3421 actgctgaac agcaaaaagt cagatagcac cacatagcag acccgccata aaacgccctg
     3481 agaagcccgt gacgggcttt tcttgtatta tgggtagttt ccttgcatga atccataaaa
     3541 ggcgcctgta gtgccattta cccccattca ctgccagagc cgtgagcgca gcgaactgaa
     3601 tgtcacgaaa aagacagcga ctcaggtgcc tgatggtcgg agacaaaagg aatattcagc
     3661 gatttgcccg agcttgcgag ggtgctactt aagcctttag ggttttaagg tctgttttgt
     3721 agaggagcaa acagcgtttg cgacatcctt ttgtaatact gcggaactga ctaaagtagt
     3781 gagttataca cagggctggg atctattctt tttatctttt tttattcttt ctttattcta
     3841 taaattataa ccacttgaat ataaacaaaa aaaacacaca aaggtctagc ggaatttaca
     3901 gagggtctag cagaatttac aagttttcca gcaaaggtct agcagaattt acagataccc
     3961 acaactcaaa ggaaaaggac tagtaattat cattgactag cccatctcaa ttggtatagt
     4021 gattaaaatc acctagacca attgagatgt atgtctgaat tagttgtttt caaagcaaat
     4081 gaactagcga ttagtcgcta tgacttaacg gagcatgaaa ccaagctaat tttatgctgt
     4141 gtggcactac tcaaccccac gattgaaaac cctacaagga aagaacggac ggtatcgttc
     4201 acttataacc aatacgctca gatgatgaac atcagtaggg aaaatgctta tggtgtatta
     4261 gctaaagcaa ccagagagct gatgacgaga actgtggaaa tcaggaatcc tttggttaaa
     4321 ggctttgaga ttttccagtg gacaaactat gccaagttct caagcgaaaa attagaatta
     4381 gtttttagtg aagagatatt gccttatctt ttccagttaa aaaaattcat aaaatataat
     4441 ctggaacatg ttaagtcttt tgaaaacaaa tactctatga ggatttatga gtggttatta
     4501 aaagaactaa cacaaaagaa aactcacaag gcaaatatag agattagcct tgatgaattt
     4561 aagttcatgt taatgcttga aaataactac catgagttta aaaggcttaa ccaatgggtt
     4621 ttgaaaccaa taagtaaaga tttaaacact tacagcaata tgaaattggt ggttgataag
     4681 cgaggccgcc cgactgatac gttgattttc caagttgaac tagatagaca aatggatctc
     4741 gtaaccgaac ttgagaacaa ccagataaaa atgaatggtg acaaaatacc aacaaccatt
     4801 acatcagatt cctacctacg taacggacta agaaaaacac tacacgatgc tttaactgca
     4861 aaaattcagc tcaccagttt tgaggcaaaa tttttgagtg acatgcaaag taagcatgat
     4921 ctcaatggtt cgttctcatg gctcacgcaa aaacaacgaa ccacactaga gaacatactg
     4981 gctaaatacg gaaggatctg aggttcttat ggctcttgta tctatcagtg aagcatcaag
     5041 actaacaaac aaaagtagaa caactgttca ccgttagata tcaaagggaa aactgtccat
     5101 atgcacagat gaaaacggtg taaaaaagat agatacatca gagcttttac gagtttttgg
     5161 tgcatttaaa gctgttcacc atgaacagat cgacaatgta acagatgaac agcatgtaac
     5221 acctaataga acaggtgaaa ccagtaaaac aaagcaacta gaacatgaaa ttgaacacct
     5281 gagacaactt gttacagctc aacagtcaca catagacagc ctgaaacagg cgatgctgct
     5341 tatcgaatca aagctgccga caacacggga gccagtgacg cctcccgtgg ggaaaaaatc
     5401 atggcaattc tggaagaaat agcgctttca gccggcaaac ctgaagccgg atctgcgatt
     5461 ctgataacaa actagcaaca ccagaacagc ccgtttgcgg gcagcaaaac ccgtacccta
     5521 ggtctagggc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
     5581 acgaaaggcc cagtctttcg actgagcctt tcgttttatt tgatgcctct ag
//
