LOCUS       Exported                5630 bp ds-DNA     circular SYN 27-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSC101-GFPmut2-mScarlet-I A4
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5630)
  AUTHORS   Mariliis
  TITLE     Direct Submission
  JOURNAL   Exported Monday, August 28, 2023 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5630
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     CDS             13..712
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /note="mScarlet-I"
                     /translation="MVSKGKRGSDQGVHAVQGAHGGLHERPRVRDRGRGRGPPLRGHPD
                     RQAEGDQGWPPALLLGHPVPSVHVRLQGLHQAPRRHPRLL*AVLPRGLQVGARDELRGR
                     RRRDRDPGHLPGGRHPDLQGEAPRHQLPS*RPRNAEEDNGLGSVHRAVVPRGRRAEGRH
                     *DGPAPEGRRPLPGGLQDHLQGQEARADARRLQRRPQVGHHLPQRGLHRGGTVRTLRGP
                     PLHRRHGRAVQ"
     CDS             815..838
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /note="8xHis"
                     /translation="HHHHHHHH"
     misc_feature    872..873
                     /note="trnscr_start"
                     /note="Tac promoter transcription start site"
     terminator      923..1017
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     rep_origin      1509..1731
                     /note="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             1779..2729
                     /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(3264..3350)
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3556..4416)
                     /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(4417..4521)
                     /gene="bla"
                     /note="AmpR promoter"
     misc_feature    4573..4600
                     /note="rrnB T2 terminator"
     misc_feature    4721..4726
                     /note="-35"
     misc_feature    4740..4745
                     /note="-10"
     misc_feature    4755
                     /note="dnaKp1"
                     /note="transcription start of promoter P1 (rpoH dependent)"
     misc_feature    4806..4871
                     /note="Tet_prom_to_SD"
     promoter        4808..4836
                     /note="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             4900..5625
                     /codon_start=1
                     /product="GFP"
                     /note="GFPmut2"
                     /translation="MKGSSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYIMADKQKDG
                     IKVNFKIRHNIEDGSVQLADHYQRNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITHGMDELYK"
     CDS             4912..4919
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /note="yeGFP"
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SK"
ORIGIN
        1 aggaggaaca atatggtgag caagggcaaa agaggcagtg atcaaggagt tcatgcggtt
       61 caaggtgcac atggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga
      121 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct
      181 gcccttctcc tgggacatcc tgtcccctca gttcatgtac ggctccaggg ccttcatcaa
      241 gcaccccgcc gacatccccg actactataa gcagtccttc cccgagggct tcaagtggga
      301 gcgcgtgatg aacttcgagg acggcggcgc cgtgaccgtg acccaggaca cctccctgga
      361 ggacggcacc ctgatctaca aggtgaagct ccgcggcacc aacttccctc ctgacggccc
      421 cgtaatgcag aagaagacaa tgggctggga agcgtccacc gagcggttgt accccgagga
      481 cggcgtgctg aagggcgaca ttaagatggc cctgcgcctg aaggacggcg gccgctacct
      541 ggcggacttc aagaccacct acaaggccaa gaagcccgtg cagatgcccg gcgcctacaa
      601 cgtcgaccgc aagttggaca tcacctccca caacgaggac tacaccgtgg tggaacagta
      661 cgaacgctcc gagggccgcc actccaccgg cggcatggac gagctgtaca agtgaccgcc
      721 taatgagcgg gctttaatca ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga
      781 gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc accaccacta
      841 attgattaat acctaggctg ctaaacaaag cccatggatc gatagctggt cgacaggact
      901 ctggggttcg agagctcgct tggactcctg ttgatagatc cagtaatgac ctcagaactc
      961 catctggatt tgttcagaac gctcggttgc cgccgggcgt tttttattgg tgagaatcca
     1021 agcactaggg acagtaagac gggtaagcct gttgatgata ccgctgcctt actgggtgca
     1081 ttagccagtc tgaatgacct gtcacgggat aatccgaagt ggtcagactg gaaaatcaga
     1141 gggcaggaac tgctgaacag caaaaagtca gatagcacca catagcagac ccgccataaa
     1201 acgccctgag aagcccgtga cgggcttttc ttgtattatg ggtagtttcc ttgcatgaat
     1261 ccataaaagg cgcctgtagt gccatttacc cccattcact gccagagccg tgagcgcagc
     1321 gaactgaatg tcacgaaaaa gacagcgact caggtgcctg atggtcggag acaaaaggaa
     1381 tattcagcga tttgcccgag cttgcgaggg tgctacttaa gcctttaggg ttttaaggtc
     1441 tgttttgtag aggagcaaac agcgtttgcg acatcctttt gtaatactgc ggaactgact
     1501 aaagtagtga gttatacaca gggctgggat ctattctttt tatctttttt tattctttct
     1561 ttattctata aattataacc acttgaatat aaacaaaaaa aacacacaaa ggtctagcgg
     1621 aatttacaga gggtctagca gaatttacaa gttttccagc aaaggtctag cagaatttac
     1681 agatacccac aactcaaagg aaaaggacta gtaattatca ttgactagcc catctcaatt
     1741 ggtatagtga ttaaaatcac ctagaccaat tgagatgtat gtctgaatta gttgttttca
     1801 aagcaaatga actagcgatt agtcgctatg acttaacgga gcatgaaacc aagctaattt
     1861 tatgctgtgt ggcactactc aaccccacga ttgaaaaccc tacaaggaaa gaacggacgg
     1921 tatcgttcac ttataaccaa tacgctcaga tgatgaacat cagtagggaa aatgcttatg
     1981 gtgtattagc taaagcaacc agagagctga tgacgagaac tgtggaaatc aggaatcctt
     2041 tggttaaagg ctttgagatt ttccagtgga caaactatgc caagttctca agcgaaaaat
     2101 tagaattagt ttttagtgaa gagatattgc cttatctttt ccagttaaaa aaattcataa
     2161 aatataatct ggaacatgtt aagtcttttg aaaacaaata ctctatgagg atttatgagt
     2221 ggttattaaa agaactaaca caaaagaaaa ctcacaaggc aaatatagag attagccttg
     2281 atgaatttaa gttcatgtta atgcttgaaa ataactacca tgagtttaaa aggcttaacc
     2341 aatgggtttt gaaaccaata agtaaagatt taaacactta cagcaatatg aaattggtgg
     2401 ttgataagcg aggccgcccg actgatacgt tgattttcca agttgaacta gatagacaaa
     2461 tggatctcgt aaccgaactt gagaacaacc agataaaaat gaatggtgac aaaataccaa
     2521 caaccattac atcagattcc tacctacgta acggactaag aaaaacacta cacgatgctt
     2581 taactgcaaa aattcagctc accagttttg aggcaaaatt tttgagtgac atgcaaagta
     2641 agcatgatct caatggttcg ttctcatggc tcacgcaaaa acaacgaacc acactagaga
     2701 acatactggc taaatacgga aggatctgag gttcttatgg ctcttgtatc tatcagtgaa
     2761 gcatcaagac taacaaacaa aagtagaaca actgttcacc gttagatatc aaagggaaaa
     2821 ctgtccatat gcacagatga aaacggtgta aaaaagatag atacatcaga gcttttacga
     2881 gtttttggtg catttaaagc tgttcaccat gaacagatcg acaatgtaac agatgaacag
     2941 catgtaacac ctaatagaac aggtgaaacc agtaaaacaa agcaactaga acatgaaatt
     3001 gaacacctga gacaacttgt tacagctcaa cagtcacaca tagacagcct gaaacaggcg
     3061 atgctgctta tcgaatcaaa gctgccgaca acacgggagc cagtgacgcc tcccgtgggg
     3121 aaaaaatcat ggcaattctg gaagaaatag cgctttcagc cggcaaacct gaagccggat
     3181 ctgcgattct gataacaaac tagcaacacc agaacagccc gtttgcgggc agcaaaaccc
     3241 gtaccctagg tctagggcgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
     3301 cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctctag
     3361 agcttgcatg cccaaggaga tggcgcccaa cagtcccccg gccacggggc ctgccaccat
     3421 acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt ccccatcggt
     3481 gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc cggccacgat
     3541 gcgtccggcg tagagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3601 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3661 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3721 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3781 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3841 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     3901 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3961 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4021 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4081 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4141 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     4201 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4261 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4321 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4381 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4441 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4501 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4561 cattattatc ataaaaggcc atccgtcagg atggccttct atgaatgcct tggctgcgat
     4621 tcattcttta tatgaataaa attgctgtca attttacgtc ttgtcctgcc atatcgcgaa
     4681 atttctgcgc aaaagcacaa aaaatttttg catctccccc ttgatgacgt ggtttacgac
     4741 cccatttagt agtcaaccgc agtgagtgag tctgacgagg ccctttcgtc ttcaagaatt
     4801 ctcatgtttg acagcttatc atcgataagc tttaatgcgg tagtttatca cagttaaatt
     4861 gctaacgcag tccggggaat tcaggcttgg aggaacaata tgaaaggatc ctctaaaggt
     4921 gaagaattat tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg
     4981 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt
     5041 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc
     5101 gcgtatggtc ttcaatgctt tgcgagatac ccagatcata tgaaacagca tgactttttc
     5161 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg
     5221 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag
     5281 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac
     5341 tataactcac acaaagtata catcatggca gacaaacaaa aggatggaat caaagttaac
     5401 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacga
     5461 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     5521 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     5581 acagctgctg ggattacaca tggtatggat gaattgtaca aataaccgcc
//
