LOCUS       Exported                6167 bp ds-DNA     circular SYN 11-SEP-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6167)
  AUTHORS   M. Tominaga et al
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Oct 7, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6167
                     /organism="synthetic DNA construct"
                     /lab_host="Pichia pastoris"
                     /mol_type="other DNA"
     source          2255..2747
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label=source:synthetic DNA construct
                     /note="/ApEinfo_hidden"
                     /note="/locus_tag=source:synthetic DNA construct"
                     /note="color: #00df87"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                     1:  201 ..  269 / #ccffcc / signal sequence
                     2:  270 .. 1061 / #ccffcc
                     Cleavage site after base 269"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori (ColE1)
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /note="color: #31849b"
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This forward directional feature has 3 segments:
                     1: 2144 .. 2149 / #ffffff / -35
                     2: 2150 .. 2167 / #ffffff
                     3: 2168 .. 2174 / #ffffff / -10"
     protein_bind    2182..2198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
                     /note="color: #31849b"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     2234..2285
                     /label=MT2029
                     /note="color: 黒; sequence: 
                     cAAGCTTGCGGCCGCGGATCcatcgatttgtatgtgaaatagctgaaattcg; 
                     added: 2024-08-31"
     terminator      2255..2747
                     /locus_tag="PpGAPDHt"
                     /label=KpGAPDH terminator
                     /label=PpGAPDHt
                     /note="color: #7eff74"
     primer_bind     complement(2722..2768)
                     /label=MT2030
                     /note="color: 黒; sequence: 
                     ttcgtaTCATACATAggatcccatgttcaattatagatccacgagtg; added: 
                     2024-08-31"
     promoter        2754..2990
                     /label=KpPhl1.1
                     /note="This forward directional feature has 6 segments:
                     1: 2754 .. 2758 / #------
                     2: 2759 .. 2788 / #99ccff / phlO
                     3: 2789 .. 2793 / #------
                     4: 2794 .. 2828 / #666699 / AOX1 promoter (-197_-163)
                     5: 2829 .. 2837 / #ff0000 / TATA box
                     6: 2838 .. 2990 / #666699 / AOX1 promoter (-153_-1)"
     CDS             2997..3713
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="This feature has 3 segments:
                     1: 2997 .. 2999 / #05fc14
                     2: 3000 .. 3002 / #05fc14 / 1a
                     3: 3003 .. 3713 / #05fc14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     terminator      3791..4037
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    4069..4668
                     /locus_tag="T38473"
                     /label=T38473
                     /note="color: #72f525"
     CDS             complement(4686..5480)
                     /codon_start=1
                     /label=KanMX
                     /note="color: #993366"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(5481..5766)
                     /label=GAP promoter (Ogataea parapolymorpha)
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(5773..5789)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gcggccgcgg atccatcgat ttgtatgtga aatagctgaa
     2281 attcgaaaat ttcattatgg ctgtatctac tttagcgtat taggcatttg agcattggct
     2341 tgaacaatgc gggctgtagt gtgtcaccaa agaaaccatt cgggttcgga tctggaagtc
     2401 ctcatcacgt gatgccgatc tcgtgtattt tattttcaga taacacctga agacttttgg
     2461 gtcggaggac tggctctttc cgatcaaatt ggaatggaaa attgctcctc taagaaaggg
     2521 tgccaacact ctttgtaaca caggacaccg tttattgcta actcgattgc attctttcct
     2581 ttcccacacc gggatctggt cttggtgaac aatctctcct gtccttatct aaatatatca
     2641 tcgcactgta accttcctta ttacttttcg agcgtccgtc ctgtattatc ttcaacctga
     2701 aaccaaactc taaccaggct tcactcgtgg atctataatt gaacatggga tcctatgtat
     2761 gatacgaaac gtaccgtatc gttaaggtag cgtcaaaatt taactgttct aacccctact
     2821 tgacagcaat atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca
     2881 ttattagctt actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga
     2941 cttttaacga caacttgaga agatcaaaaa acaactaatt attcgaaacg actagtatgg
     3001 tttctaaggg tgaagagttg ttcactggtg ttgttccaat cttggttgag ttggacggtg
     3061 acgttaacgg acacaagttc tctgtttctg gtgaaggtga gggtgacgct acttacggaa
     3121 agttgacttt gaagttcatc tgtactactg gtaagttgcc agttccatgg ccaactttgg
     3181 ttactacttt gacttacggt gttcagtgtt tctccagata cccagaccac atgaagcagc
     3241 acgatttctt caagtctgct atgccagagg gttacgttca agagagaact atcttcttca
     3301 aggacgacgg taactacaag actagagctg aggttaagtt cgagggtgac acattggtta
     3361 acagaatcga gttgaagggt atcgacttca aagaggacgg aaacatcttg ggtcacaagt
     3421 tggagtacaa ctacaactcc cacaacgttt acatcatggc tgacaagcag aagaacggta
     3481 tcaaggttaa cttcaagatc agacacaaca tcgaggacgg ttccgttcaa ttggctgacc
     3541 actaccaaca gaacactcca attggtgacg gtccagtttt gttgccagac aaccactact
     3601 tgtccactca atccgctttg tccaaggacc caaacgagaa gagagatcac atggttttgt
     3661 tggagttcgt tactgctgct ggtatcactt tgggtatgga cgagttgtac aagtaactcg
     3721 agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc
     3781 tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt
     3841 tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc
     3901 tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg
     3961 tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta
     4021 cagaagatta agtgagacgt tcgtttgtgc aagcttagat cttctagaga caataagaag
     4081 aaaaaaaaag aaaagcggtg ggggagggat tattaaataa ggattatgta accccagggt
     4141 accgttctat acatatttaa ggattattta ggacaatcga tgaaatcggc atcaaactgg
     4201 atgggagtat agtgtccgga taatcggata aatcatcttg cgaggagccg cttggttggt
     4261 tggtgagagg agtgaaatat gtgtctcctc acccaagaat cgcgatatca gcaccctgtg
     4321 ggggacacta ttggcctccc tcccaaacct tcgatgtggt agtgctttat tatattgatt
     4381 acattgatta catagctaaa ccctgcctgg ttgcaagttg agctccgaat tccaatatta
     4441 gtaaaatgcc tgcaagataa cctcggtatg gcgtccgacc ccgcttaatt attttaactc
     4501 ctttccaacg aggacttcgt aatttttgat tagggagttg agaaacgggg ggtcttgata
     4561 cctcctcgat ttcagatccc accccctctc agtcccaagt gggacccccc tcggccgtga
     4621 aatgcgcgca ctttagtttt tttcgcatgt aaacgccggt gtccgtcatc tagagaattc
     4681 cccgctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
     4741 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
     4801 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
     4861 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
     4921 gtcacgacga gatcctcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
     4981 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
     5041 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
     5101 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
     5161 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
     5221 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
     5281 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
     5341 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
     5401 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
     5461 aatccatctt gttcaatcat atggtttcta tattatcttt gtactaaaga gcaattgata
     5521 atgtgcgaga aaaactggtc cttatatacc gtttgcagca ctccctcccg aactttacga
     5581 aaagtcgtgc gccacctgat tttcatcacg ccaaaaacct acacgtatga ctactccggg
     5641 ccagtgttca ccacgagcta tatagtgtta attaattacc ttattggtta gctctgcatg
     5701 taagggtggt gtgagccggg aattgggtct actctagcgt tcagtaaggt gatataaagc
     5761 tctgtagaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
     5821 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
     5881 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
     5941 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     6001 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
     6061 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     6121 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcga
//
