LOCUS       Exported                5626 bp ds-DNA     circular SYN 28-AUG-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSC101-GFPmut2-mScarlet-I
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5626)
  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..5626
                     /organism="synthetic DNA construct"
                     /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..2978
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /note="mScarlet-I"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     CDS             3081..3104
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /note="8xHis"
                     /translation="HHHHHHHH"
     misc_feature    3138..3139
                     /note="trnscr_start"
                     /note="Tac promoter transcription start site"
     terminator      3189..3283
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     rep_origin      3775..3997
                     /note="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4045..4995
                     /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(5530..5616)
                     /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 caagggcgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg
     2341 agggctccat gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg
     2401 agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttctcctggg
     2461 acatcctgtc ccctcagttc atgtacggct ccagggcctt catcaagcac cccgccgaca
     2521 tccccgacta ctataagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact
     2581 tcgaggacgg cggcgccgtg accgtgaccc aggacacctc cctggaggac ggcaccctga
     2641 tctacaaggt gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga
     2701 agacaatggg ctgggaagcg tccaccgagc ggttgtaccc cgaggacggc gtgctgaagg
     2761 gcgacattaa gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga
     2821 ccacctacaa ggccaagaag cccgtgcaga tgcccggcgc ctacaacgtc gaccgcaagt
     2881 tggacatcac ctcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgagg
     2941 gccgccactc caccggcggc atggacgagc tgtacaagtg accgcctaat gagcgggctt
     3001 taatcactag tgaattcgcg gccgcctgca ggtcgaccat atgggagagc tccgtcgaca
     3061 agcttgcggc cgcactcgag caccaccacc accaccacca ccactaattg attaatacct
     3121 aggctgctaa acaaagccca tggatcgata gctggtcgac aggactctgg ggttcgagag
     3181 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     3241 cagaacgctc ggttgccgcc gggcgttttt tattggtgag aatccaagca ctagggacag
     3301 taagacgggt aagcctgttg atgataccgc tgccttactg ggtgcattag ccagtctgaa
     3361 tgacctgtca cgggataatc cgaagtggtc agactggaaa atcagagggc aggaactgct
     3421 gaacagcaaa aagtcagata gcaccacata gcagacccgc cataaaacgc cctgagaagc
     3481 ccgtgacggg cttttcttgt attatgggta gtttccttgc atgaatccat aaaaggcgcc
     3541 tgtagtgcca tttaccccca ttcactgcca gagccgtgag cgcagcgaac tgaatgtcac
     3601 gaaaaagaca gcgactcagg tgcctgatgg tcggagacaa aaggaatatt cagcgatttg
     3661 cccgagcttg cgagggtgct acttaagcct ttagggtttt aaggtctgtt ttgtagagga
     3721 gcaaacagcg tttgcgacat ccttttgtaa tactgcggaa ctgactaaag tagtgagtta
     3781 tacacagggc tgggatctat tctttttatc tttttttatt ctttctttat tctataaatt
     3841 ataaccactt gaatataaac aaaaaaaaca cacaaaggtc tagcggaatt tacagagggt
     3901 ctagcagaat ttacaagttt tccagcaaag gtctagcaga atttacagat acccacaact
     3961 caaaggaaaa ggactagtaa ttatcattga ctagcccatc tcaattggta tagtgattaa
     4021 aatcacctag accaattgag atgtatgtct gaattagttg ttttcaaagc aaatgaacta
     4081 gcgattagtc gctatgactt aacggagcat gaaaccaagc taattttatg ctgtgtggca
     4141 ctactcaacc ccacgattga aaaccctaca aggaaagaac ggacggtatc gttcacttat
     4201 aaccaatacg ctcagatgat gaacatcagt agggaaaatg cttatggtgt attagctaaa
     4261 gcaaccagag agctgatgac gagaactgtg gaaatcagga atcctttggt taaaggcttt
     4321 gagattttcc agtggacaaa ctatgccaag ttctcaagcg aaaaattaga attagttttt
     4381 agtgaagaga tattgcctta tcttttccag ttaaaaaaat tcataaaata taatctggaa
     4441 catgttaagt cttttgaaaa caaatactct atgaggattt atgagtggtt attaaaagaa
     4501 ctaacacaaa agaaaactca caaggcaaat atagagatta gccttgatga atttaagttc
     4561 atgttaatgc ttgaaaataa ctaccatgag tttaaaaggc ttaaccaatg ggttttgaaa
     4621 ccaataagta aagatttaaa cacttacagc aatatgaaat tggtggttga taagcgaggc
     4681 cgcccgactg atacgttgat tttccaagtt gaactagata gacaaatgga tctcgtaacc
     4741 gaacttgaga acaaccagat aaaaatgaat ggtgacaaaa taccaacaac cattacatca
     4801 gattcctacc tacgtaacgg actaagaaaa acactacacg atgctttaac tgcaaaaatt
     4861 cagctcacca gttttgaggc aaaatttttg agtgacatgc aaagtaagca tgatctcaat
     4921 ggttcgttct catggctcac gcaaaaacaa cgaaccacac tagagaacat actggctaaa
     4981 tacggaagga tctgaggttc ttatggctct tgtatctatc agtgaagcat caagactaac
     5041 aaacaaaagt agaacaactg ttcaccgtta gatatcaaag ggaaaactgt ccatatgcac
     5101 agatgaaaac ggtgtaaaaa agatagatac atcagagctt ttacgagttt ttggtgcatt
     5161 taaagctgtt caccatgaac agatcgacaa tgtaacagat gaacagcatg taacacctaa
     5221 tagaacaggt gaaaccagta aaacaaagca actagaacat gaaattgaac acctgagaca
     5281 acttgttaca gctcaacagt cacacataga cagcctgaaa caggcgatgc tgcttatcga
     5341 atcaaagctg ccgacaacac gggagccagt gacgcctccc gtggggaaaa aatcatggca
     5401 attctggaag aaatagcgct ttcagccggc aaacctgaag ccggatctgc gattctgata
     5461 acaaactagc aacaccagaa cagcccgttt gcgggcagca aaacccgtac cctaggtcta
     5521 gggcggcgga tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa
     5581 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctctag
//
