LOCUS       62056_3205        4641 bp DNA     circular SYN 10-NOV-2020
DEFINITION  Cloning vector pAS_HO_natNT2, complete sequence.
ACCESSION   MT684463
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4641)
  AUTHORS   Cromie G.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-JUN-2020) Dudley Lab, PNRI, 720 Broadway, Seattle, WA 
            98027, USA
REFERENCE   2  (bases 1 to 4641)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (30-JUN-2020) Dudley Lab, PNRI, 720 Broadway, Seattle, WA 98027, 
            USA"
FEATURES             Location/Qualifiers
     source          1..4641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(719..823)
                     /label=AmpR promoter
     primer_bind     1297..1313
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1749..2092
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      2676..2863
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3338..3354)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3362..3378)
                     /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(3386..3416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3431..3452)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3740..4328)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4502..4641,1..718))
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
       61 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
      121 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
      181 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
      241 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
      301 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
      361 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
      421 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
      481 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
      541 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
      601 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
      661 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
      721 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
      781 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      841 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
      901 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
      961 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     1021 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     1081 agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     1141 aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     1201 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     1261 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcg
     1321 agctcggtac ccggggcggc cgccgtcacg gctaactctt acgttatgtg cgcagatggc
     1381 tccgctgccc gcgtcataaa tgtcacacag ggctatcaga aaatctataa tatacagcaa
     1441 aaaaccaaac acagagcttt tgaaggtgaa cctggtaggt tagatcccag gcgtagaaca
     1501 gtttatcagc gtcttgcatt acaatgtact gcaggtcata aattgtcagt cagggtccct
     1561 accaaaccac tgttggaaaa aagtggtaga aatgccacca aatataaagt gagatggaga
     1621 aatctgcagc aatgtcagac gcttgatggt agagcgctcg tacgctgcag gtcgacggat
     1681 ccccgggtta attaaggcgc gccagatctg tttagcttgc ctcgtccccg ccgggtcacc
     1741 cggccagcga catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag
     1801 gggcatgatg tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg
     1861 catccataca ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga
     1921 cctgcgagca gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc
     1981 tgtagagaaa tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt
     2041 ttaaaatctt gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtac
     2101 cactcttgac gacacggctt accggtaccg caccagtgtc ccgggggacg ccgaggccat
     2161 cgaggcactg gatgggtcct tcaccaccga caccgtcttc cgcgtcaccg ccaccgggga
     2221 cggcttcacc ctgcgggagg tgccggtgga cccgcccctg accaaggtgt tccccgacga
     2281 cgaatcggac gacgaatcgg acgacgggga ggacggcgac ccggactccc ggacgttcgt
     2341 cgcgtacggg gacgacggcg acctggcggg cttcgtggtc atctcgtact cggcgtggaa
     2401 ccgccggctg accgtcgagg acatcgaggt cgccccggag caccgggggc acggggtcgg
     2461 gcgcgcgttg atggggctcg cgacggagtt cgccggcgag cggggcgccg ggcacctctg
     2521 gctggaggtc accaacgtca acgcaccggc gatccacgcg taccggcgga tggggttcac
     2581 cctctgcggc ctggacaccg ccctgtacga cggcaccgcc tcggacggcg agcggcaggc
     2641 gctctacatg agcatgccct gcccctaatc tcgaggcgaa tttcttatga tttatgattt
     2701 ttattattaa ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg
     2761 ttttaaaacg aaaattcttg ttcttgagta actctttcct gtaggtcagg ttgctttctc
     2821 aggtatagca tgaggtcgct cttattgacc acacctctac cggcatgccg agcaaatgcc
     2881 tgcaaatcgc tccccatttc acccaattgt agatatgcta actccagcaa tgagttgatg
     2941 aatctcggtg tgtattttat gtcctcagag gacaacacct gttgtaatcg agctcgaatt
     3001 catcgatgca ctgtgttgct tgtcccagaa agggaatcaa gtatttttat aaagattgga
     3061 gtggtaaaaa tcgagtatgt gctagatgct atggaagata caaattcagc ggtcatcact
     3121 gtataaattg caagtatgta ccagaagcac gtgaagtgaa aaaggcaaaa gacaaaggcg
     3181 aaaaattggg cattacgccc gaaggtttgc cagttaaagg accagagtgt ataaaatgtg
     3241 gcggaatctt acagtttgat gctgtccgcg ggcctcataa gagttgtggt aacaacgcag
     3301 gcggccgctg caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     3361 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     3421 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     3481 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3541 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3601 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3661 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3721 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3781 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3841 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     3901 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     3961 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4021 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4081 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4141 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4201 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4261 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4321 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4381 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4441 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4501 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4561 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4621 gtgctgcaat gataccgcga g
//
