LOCUS       sequence-41672-e        3576 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3576)
  AUTHORS   Park HS, Hohn MJ, Umehara T, Guo LT, Osborne EM, Benner J, Noren CJ,
            Rinehart J, Soll D
  TITLE     Expanding the genetic code of Escherichia coli with phosphoserine.
  JOURNAL   Science. 2011 Aug 26;333(6046):1151-4.
  PUBMED    21868676
REFERENCE   2  (bases 1 to 3576)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science."; 
            date: "2011-08-26"; volume: "333(6046)"; pages: "1151-4"
COMMENT     Sequence Label: pMYO127TAG-SepT
FEATURES             Location/Qualifiers
     source          1..3576
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(277..294)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(277..293)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(286..308)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        complement(join(513..518,519..535,536..541))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     rep_origin      653..1198
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     1099..1118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1315..1332
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        1724..1826
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1827..2486
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1893..1912)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3210..3227
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3258..3344
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3436..3463
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgccgcttct ttgagcgaac
       61 gatcaaaaat aagtggcgcc ccatcaaaaa aatattctca acataaaaaa ctttgtgtaa
      121 tacttgtaac gctgaattcg ccggggtagt ctaggggtta ggcagcggac tctagatccg
      181 ccttacgtgg gttcaaatcc cacccccggc tccaggatcc tctagagtcg acctgcagat
      241 ccttagcgaa agctaaggat tttttttaag cttggcactg gccgtcgttt tacaacgtcg
      301 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      361 cagatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag
      421 ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact
      481 gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca
      541 aacatgagaa ttacaactta tatcgtatgg ggctgacttc aggtgctaca tttgaagaga
      601 taaattgcac tgaaatctag aaatatttta tctgattaat aagatgatct tcttgagatc
      661 gttttggtct gcgcgtaatc tcttgctctg aaaacgaaaa aaccgccttg cagggcggtt
      721 tttcgaaggt tctctgagct accaactctt tgaaccgagg taactggctt ggaggagcgc
      781 agccaccaaa acttgtcctt tcagtttagc cttaaccggc gcatgacttc aagactaact
      841 cctctaaatc aattaccagt ggctgctgcc agtggtgctt ttgcatgtct ttccgggttg
      901 gactcaagac gatagttacc ggataaggcg cagcggtcgg actgaacggg gggttcgtgc
      961 atacagtcca gcttggagcg aactgcctac ccggaactga gtgtcaggcg tggaatgaga
     1021 caaacgcggc cataacagcg gaatgacacc ggtaaaccga aaggcaggaa caggagagcg
     1081 cacgagggag ccgccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     1141 ccactgattt gagcgtcaga tttcgtgatg cttgtcaggg gggcggagcc tatggaaaaa
     1201 cggctttgcc gcggccctct cacttccctg ttaagtatct tcctggcatc ttccaggaaa
     1261 tctccgcccc gttcgtaagc catttccgct cgccgcagtc gaacgaccga gcgtagcgag
     1321 tcagtgagcg aggaagcgga atatatcctg tatcacatat tctgctgacg caccggtgca
     1381 gccttttttc tcctgccaca tgaagcactt cactgacacc ctcatcagtg ccaacatagt
     1441 aagccagtat acactccgct agcgctgatg tccggcggtg cttttgccgt tacgcaccac
     1501 cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg gagcacctca
     1561 aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca ctttgcgccg
     1621 aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt gtccctgttg
     1681 ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg cacgtaagag
     1741 gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt gagttatcga
     1801 gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat accaccgttg
     1861 atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta
     1921 cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta aagaaaaata
     1981 agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat gctcatccgg
     2041 aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt cacccttgtt
     2101 acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa taccacgacg
     2161 atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt gaaaacctgg
     2221 cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat ccctgggtga
     2281 gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc cccgttttca
     2341 ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg attcaggttc
     2401 atcatgccgt ctgtgatggc ttccatgtcg gcagaatgct taatgaatta caacagtact
     2461 gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc ccttaaacgc
     2521 ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat tcgaaagcaa
     2581 attcgacccg ggcggccgca acccagcgtt cgatgcttct ttgagcgaac gatcaaaaat
     2641 aagtgccttc ccatcaaaaa aatattctca acataaaaaa ctttgtgtaa tacttgtaac
     2701 gctatatgga gattaactca atctagaggg tattacatat ggttctgtct gaaggtgaat
     2761 ggcagctggt tctgcacgtt tgggctaaag ttgaagctga cgttgctggt cacggtcagg
     2821 acatcctgat ccgtctgttc aaatctcacc cggaaaccct ggaaaaattc gaccgtttca
     2881 aacacctgaa aaccgaagct gaaatgaagg cttctgaaga cctgaaaaaa cacggtgtta
     2941 ccgttctgac cgctctgggt gctatcctga agaaaaaggg tcaccacgaa gctgaactga
     3001 aaccgctggc tcagtctcac gctaccaaac acaaaatccc gatcaaatac ctggagttca
     3061 tctctgaagc tatcatccac gttctgcact ctcgtcatcc gggtaacttc ggtgcttagg
     3121 ctcagggtgc tatgaacaaa gctctggaac tgttccgtaa agacatcgct gctaaataca
     3181 aagaactggg ttaccagggt ggttctggtc atcaccatca ccatcactaa ggcgcgccgt
     3241 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     3301 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
     3361 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
     3421 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
     3481 ttttgagatc tatgccccgc gcccaccgga aggagctacc ggcagcggtg cggactgttg
     3541 taactcagaa taagaaatga ggccgctcat ggcgtt
//
