LOCUS       Exported                5633 bp ds-DNA     circular SYN 08-NOV-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSC101-GFPmut2-mScarlet-I A6T
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5633)
  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..5633
                     /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..2985
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /note="mScarlet-I"
                     /translation="MVSKGKK*GSDQGVHAVQGAHGGLHERPRVRDRGRGRGPPLRGHP
                     DRQAEGDQGWPPALLLGHPVPSVHVRLQGLHQAPRRHPRLL*AVLPRGLQVGARDELRG
                     RRRRDRDPGHLPGGRHPDLQGEAPRHQLPS*RPRNAEEDNGLGSVHRAVVPRGRRAEGR
                     H*DGPAPEGRRPLPGGLQDHLQGQEARADARRLQRRPQVGHHLPQRGLHRGGTVRTLRG
                     PPLHRRHGRAVQ"
     CDS             3088..3111
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /note="8xHis"
                     /translation="HHHHHHHH"
     misc_feature    3145..3146
                     /note="trnscr_start"
                     /note="Tac promoter transcription start site"
     terminator      3196..3290
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     rep_origin      3782..4004
                     /note="pSC101 ori"
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4052..5002
                     /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(5537..5623)
                     /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 caagggcaaa aaatgaggca gtgatcaagg agttcatgcg gttcaaggtg
     2341 cacatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     2401 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
     2461 tcctgggaca tcctgtcccc tcagttcatg tacggctcca gggccttcat caagcacccc
     2521 gccgacatcc ccgactacta taagcagtcc ttccccgagg gcttcaagtg ggagcgcgtg
     2581 atgaacttcg aggacggcgg cgccgtgacc gtgacccagg acacctccct ggaggacggc
     2641 accctgatct acaaggtgaa gctccgcggc accaacttcc ctcctgacgg ccccgtaatg
     2701 cagaagaaga caatgggctg ggaagcgtcc accgagcggt tgtaccccga ggacggcgtg
     2761 ctgaagggcg acattaagat ggccctgcgc ctgaaggacg gcggccgcta cctggcggac
     2821 ttcaagacca cctacaaggc caagaagccc gtgcagatgc ccggcgccta caacgtcgac
     2881 cgcaagttgg acatcacctc ccacaacgag gactacaccg tggtggaaca gtacgaacgc
     2941 tccgagggcc gccactccac cggcggcatg gacgagctgt acaagtgacc gcctaatgag
     3001 cgggctttaa tcactagtga attcgcggcc gcctgcaggt cgaccatatg ggagagctcc
     3061 gtcgacaagc ttgcggccgc actcgagcac caccaccacc accaccacca ctaattgatt
     3121 aatacctagg ctgctaaaca aagcccatgg atcgatagct ggtcgacagg actctggggt
     3181 tcgagagctc gcttggactc ctgttgatag atccagtaat gacctcagaa ctccatctgg
     3241 atttgttcag aacgctcggt tgccgccggg cgttttttat tggtgagaat ccaagcacta
     3301 gggacagtaa gacgggtaag cctgttgatg ataccgctgc cttactgggt gcattagcca
     3361 gtctgaatga cctgtcacgg gataatccga agtggtcaga ctggaaaatc agagggcagg
     3421 aactgctgaa cagcaaaaag tcagatagca ccacatagca gacccgccat aaaacgccct
     3481 gagaagcccg tgacgggctt ttcttgtatt atgggtagtt tccttgcatg aatccataaa
     3541 aggcgcctgt agtgccattt acccccattc actgccagag ccgtgagcgc agcgaactga
     3601 atgtcacgaa aaagacagcg actcaggtgc ctgatggtcg gagacaaaag gaatattcag
     3661 cgatttgccc gagcttgcga gggtgctact taagccttta gggttttaag gtctgttttg
     3721 tagaggagca aacagcgttt gcgacatcct tttgtaatac tgcggaactg actaaagtag
     3781 tgagttatac acagggctgg gatctattct ttttatcttt ttttattctt tctttattct
     3841 ataaattata accacttgaa tataaacaaa aaaaacacac aaaggtctag cggaatttac
     3901 agagggtcta gcagaattta caagttttcc agcaaaggtc tagcagaatt tacagatacc
     3961 cacaactcaa aggaaaagga ctagtaatta tcattgacta gcccatctca attggtatag
     4021 tgattaaaat cacctagacc aattgagatg tatgtctgaa ttagttgttt tcaaagcaaa
     4081 tgaactagcg attagtcgct atgacttaac ggagcatgaa accaagctaa ttttatgctg
     4141 tgtggcacta ctcaacccca cgattgaaaa ccctacaagg aaagaacgga cggtatcgtt
     4201 cacttataac caatacgctc agatgatgaa catcagtagg gaaaatgctt atggtgtatt
     4261 agctaaagca accagagagc tgatgacgag aactgtggaa atcaggaatc ctttggttaa
     4321 aggctttgag attttccagt ggacaaacta tgccaagttc tcaagcgaaa aattagaatt
     4381 agtttttagt gaagagatat tgccttatct tttccagtta aaaaaattca taaaatataa
     4441 tctggaacat gttaagtctt ttgaaaacaa atactctatg aggatttatg agtggttatt
     4501 aaaagaacta acacaaaaga aaactcacaa ggcaaatata gagattagcc ttgatgaatt
     4561 taagttcatg ttaatgcttg aaaataacta ccatgagttt aaaaggctta accaatgggt
     4621 tttgaaacca ataagtaaag atttaaacac ttacagcaat atgaaattgg tggttgataa
     4681 gcgaggccgc ccgactgata cgttgatttt ccaagttgaa ctagatagac aaatggatct
     4741 cgtaaccgaa cttgagaaca accagataaa aatgaatggt gacaaaatac caacaaccat
     4801 tacatcagat tcctacctac gtaacggact aagaaaaaca ctacacgatg ctttaactgc
     4861 aaaaattcag ctcaccagtt ttgaggcaaa atttttgagt gacatgcaaa gtaagcatga
     4921 tctcaatggt tcgttctcat ggctcacgca aaaacaacga accacactag agaacatact
     4981 ggctaaatac ggaaggatct gaggttctta tggctcttgt atctatcagt gaagcatcaa
     5041 gactaacaaa caaaagtaga acaactgttc accgttagat atcaaaggga aaactgtcca
     5101 tatgcacaga tgaaaacggt gtaaaaaaga tagatacatc agagctttta cgagtttttg
     5161 gtgcatttaa agctgttcac catgaacaga tcgacaatgt aacagatgaa cagcatgtaa
     5221 cacctaatag aacaggtgaa accagtaaaa caaagcaact agaacatgaa attgaacacc
     5281 tgagacaact tgttacagct caacagtcac acatagacag cctgaaacag gcgatgctgc
     5341 ttatcgaatc aaagctgccg acaacacggg agccagtgac gcctcccgtg gggaaaaaat
     5401 catggcaatt ctggaagaaa tagcgctttc agccggcaaa cctgaagccg gatctgcgat
     5461 tctgataaca aactagcaac accagaacag cccgtttgcg ggcagcaaaa cccgtaccct
     5521 aggtctaggg cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     5581 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctc tag
//
