LOCUS       BBF10K_000456           2411 bp    DNA     circular UNK 05-NOV-2020
DEFINITION  Pichia NTS 3'-Homology Region in pOpen-v3.
ACCESSION   BBF10K_000456
VERSION     BBF10K_000456.1
KEYWORDS    BBFD_0017; Open Yeast Collection Part; OpenMTA; FreeGenes; Golden
            Gate Assembly.
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
            .
REFERENCE   1  (bases 1 to 2001)
  AUTHORS   Keoni Gandall
  TITLE     Direct Submission
REFERENCE   2  (bases 2002 to 2411)
  AUTHORS   Scott Pownall, Open Science Network
  TITLE     Direct Submission
  JOURNAL   https://biobricks.org/freegenes/
FEATURES             Location/Qualifiers
     source          1..2001
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /original_record="pOpen_v3.1"
     primer_bind     complement(24..42)
                     /label="T7 Reverse"
                     /note="/ApEinfo_fwdcolor=#b7e6d7 /ApEinfo_revcolor=#b7e6d7"
                     /original_record="pOpen_v3.1"
     terminator      38..85
                     /label="T7 terminator"
                     /note="/ApEinfo_fwdcolor=#d6b295 /ApEinfo_revcolor=#d6b295"
                     /original_record="pOpen_v3.1"
     primer_bind     complement(86..106)
                     /label="M13 rev"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /original_record="pOpen_v3.1"
     misc_feature    107..121
                     /label="AarI"
                     /note="/ApEinfo_fwdcolor=#b1ff67 /ApEinfo_revcolor=#b1ff67"
                     /original_record="pOpen_v3.1"
     misc_feature    122..172
                     /label="spacer"
                     /note="/ApEinfo_fwdcolor=#b1ff67 /ApEinfo_revcolor=#b1ff67"
                     /original_record="pOpen_v3.1"
     rep_origin      complement(173..761)
                     /direction=LEFT
                     /label="ori"
                     /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc"
                     /original_record="pOpen_v3.1"
     CDS             complement(952..1812)
                     /label="AmpR"
                     /note="/ApEinfo_fwdcolor=#ff9ccd /ApEinfo_revcolor=#ff9ccd"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW*"
                     /original_record="pOpen_v3.1"
     misc_feature    1918..1932
                     /label="AarI"
                     /note="/ApEinfo_fwdcolor=#d6b295 /ApEinfo_revcolor=#d6b295"
                     /original_record="pOpen_v3.1"
     primer_bind     1933..1949
                     /label="M13 fwd"
                     /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc"
                     /original_record="pOpen_v3.1"
     terminator      1950..1981
                     /label="tonB terminator"
                     /note="/ApEinfo_fwdcolor=#b4abac /ApEinfo_revcolor=#b4abac"
                     /original_record="pOpen_v3.1"
     primer_bind     2033..2052
                     /label="3' Barcode"
                     /note="/ApEinfo_fwdcolor=#84b0dc /ApEinfo_revcolor=#84b0dc"
                     /original_record="part"
     misc_feature    2053..2297
                     /label="Pichia NTS 3'-Homology Region 245bp"
                     /original_record="part"
     protein_bind    2298..2305
                     /label="NotI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
                     /original_record="part"
     misc_feature    2318..2325
                     /label="random sequence"
                     /original_record="part"
     misc_feature    2002..2021
                     /label="FreeGenes dialout primer"
                     /ApEinfo_fwdcolor="#0048BA"
                     /ApEinfo_revcolor="#0048BA"
     misc_feature    complement(2332..2351)
                     /label="FreeGenes dialout primer"
                     /ApEinfo_fwdcolor="#0048BA"
                     /ApEinfo_revcolor="#0048BA"
     misc_feature    2352..2411
                     /label="FreeGenes barcode"
                     /ApEinfo_fwdcolor="#FF7E00"
                     /ApEinfo_revcolor="#FF7E00"
     protein_bind    join(2022..2027,2029..2032)
                     /label="BsaI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
     protein_bind    complement(join(2307..2310,2312..2317))
                     /label="BsaI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
     protein_bind    2373..2390
                     /label="I-SceI"
                     /ApEinfo_fwdcolor="#4F0080"
                     /ApEinfo_revcolor="#4F0080"
ORIGIN
        1 cgctgaggtg tcaatcgtcg gagccgctga gcaataacta gcataacccc ttggggcctc
       61 taaacgggtc ttgaggggtt ttttgcatgg tcatagctgt ttcctgagag cttggcaggt
      121 gatgacacac attaacaaat ttcgtgagga gtctccagaa gaatgccatt aatttccata
      181 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
      241 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
      301 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
      361 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
      421 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
      481 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
      541 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
      601 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
      661 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
      721 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaaggccta ctattagcaa
      781 caacgatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      841 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
      901 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
      961 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1021 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1081 aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc
     1141 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1201 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1261 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1321 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1381 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     1441 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     1501 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     1561 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     1621 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     1681 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     1741 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     1801 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     1921 ctgcaccagt cagtaaaacg acggccagta gtcaaaagcc tccgaccgga ggcttttgac
     1981 ttggttcagg tggagtggga gaacgatgat gctcactctc gggtctctag acagagcact
     2041 tgaatccact gctgactttt caaatattta gggtattttt ctcacttttt cctagcaaac
     2101 tccaattggt gggttcagtg caatggagta tcaccttgca accacaacgt aatagctaac
     2161 ttgtggccac catgtctggt tgtagagata attggattct aatgtggatc acatgactac
     2221 tcacgtgtca aaaacccaac ctgacttggc ccagcttagc aagaatattt cgaatccact
     2281 cttgtggcct agtggacgcg gccgcacgaa tgagaccctg acacagatat cccgggagga
     2341 agatatagca cggacaaagt cattccttcg gatagggata acagggtaat ctagctgtat
     2401 agggcgccgt c
//
