LOCUS       40924_37888        5634 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pRS413-SUP61S, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5634)
  AUTHORS   Muller HM, Annaluru N, Boeke JD, Chandrasegaran S.
  TITLE     Total Synthesis of a Functional Designer Eukaryotic Chromosome
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5634)
  AUTHORS   Muller HM, Annaluru N, Boeke JD, Chandrasegaran S.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-JUN-2013) Genomes et Genetique, Institut Pasteur, 25 
            rue du Docteur Roux, Paris 75015, France
REFERENCE   3  (bases 1 to 5634)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5634)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (10-JUN-2013) Genomes et Genetique, Institut Pasteur, 25 rue du 
            Docteur Roux, Paris 75015, France"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        317..503
                     /label=HIS3 promoter
     CDS             504..1160
                     /codon_start=1
                     /label=HIS3
                     /note="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     misc_feature    1883..1951
                     /label=LacZ alpha
                     /note="LacZ alpha"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2093..2109
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2121..2784
                     /label=synSUP61 cassette
                     /note="synSUP61 cassette"
     primer_bind     complement(2807..2823)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2860..2878)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2899..2915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2923..2939)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(2947..2977)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2992..3013)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3301..3889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4063..4920)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4921..5025)
                     /label=AmpR promoter
     misc_feature    5062..5565
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an
                     autonomously replicating sequence"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgcgtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt cccctagcgg atgactcttt ttttttctta gcgattggca ttatcacata
      421 atgaattata cattatataa agtaatgtga tttcttcgaa gaatatacta aaaaatgagc
      481 aggcaagata aacgaaggca aagatgacag agcagaaagc cctagtaaag cgtattacaa
      541 atgaaaccaa gattcagatt gcgatctctt taaagggtgg tcccctagcg atagagcact
      601 cgatcttccc agaaaaagag gcagaagcag tagcagaaca ggccacacaa tcgcaagtga
      661 ttaacgtcca cacaggtata gggtttctgg accatatgat acatgctctg gccaagcatt
      721 ccggctggtc gctaatcgtt gagtgcattg gtgacttaca catagacgac catcacacca
      781 ctgaagactg cgggattgct ctcggtcaag cttttaaaga ggccctactg gcgcgtggag
      841 taaaaaggtt tggatcagga tttgcgcctt tggatgaggc actttccaga gcggtggtag
      901 atctttcgaa caggccgtac gcagttgtcg aacttggttt gcaaagggag aaagtaggag
      961 atctctcttg cgagatgatc ccgcattttc ttgaaagctt tgcagaggct agcagaatta
     1021 ccctccacgt tgattgtctg cgaggcaaga atgatcatca ccgtagtgag agtgcgttca
     1081 aggctcttgc ggttgccata agagaagcca cctcgcccaa tggtaccaac gatgttccct
     1141 ccaccaaagg tgttcttatg tagtgacacc gattatttaa agctgcagca tacgatatat
     1201 atacatgtgt atatatgtat acctatgaat gtcagtaagt atgtatacga acagtatgat
     1261 actgaagatg acaaggtaat gcatcattct atacgtgtca ttctgaacga ggcgcgcttt
     1321 ccttttttct ttttgctttt tctttttttt tctcttgaac tcgacggatc tatgcggtgt
     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg
     2101 acggtatcga taagcttgat aactttaaat aatgccaatt atttaaagtt acaaaaagat
     2161 gagcaaaaaa acgacaccag caggatttga accagcgcgg gcagagccca agagatttcg
     2221 agtctctcgc cttaaccact cggccatagt gccttaactg atgtagtatg caaccgggga
     2281 gttagtcacg gaatggcggg ctgtctacag cggggcacag cgggcttgcg gcgactcgtc
     2341 tcgaggcagc cgcggctggg ggagtacagc agctgtatct gcaatctgca aatggagagg
     2401 cgagtaaggt atgaactatg tatattaggg agaaatgcct atggatgtgg ctggtggaca
     2461 caaaaggctg gagcgccgag ctgcagggtt tgccggccag tacagcttgc ggccgctcaa
     2521 gtatccctgg gccctgtaga gaatgcgaca gaccacagct tttcaggacg ccagtaggag
     2581 aggctgcata ggaaagcgcg cgaattgcac agctagcgct gtgcaaacta tgcgcgttgg
     2641 tgttcacgtg atgcggcgcg tataataatt agtgatcacg tgatataggt gcatatacgc
     2701 tctgcactta aaacacagca attagtttaa aacgtaaatt ctattggaaa tcgtaacttt
     2761 aaataatgcc aattatttaa agttatcgaa ttcctgcagc ccgggggatc cactagttct
     2821 agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag ggttaattgc
     2881 gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     2941 tccacacaac ataggagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     3001 gtaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     3061 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     3121 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     3181 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     3241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     3301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     3361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     3421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     3481 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     3541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3721 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     3781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3961 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4021 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     4081 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     4141 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     4201 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     4261 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     4321 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4381 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4441 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4501 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4561 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4621 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4681 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4741 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4801 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4861 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4921 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4981 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     5041 agtgccacct gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta
     5101 atataaatat ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt
     5161 tgttttccga agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt
     5221 gctcttcctg ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca
     5281 cacacgaaaa tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta
     5341 atctgctttt cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta
     5401 aacctttgtt tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc
     5461 taaaatccaa atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc
     5521 atcattaaaa gatacgaggc gcgtgtaagt tacaggcaag cgatccgtcc taagaaacca
     5581 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtc
//
