LOCUS       BBF10K_000460           2645 bp    DNA     circular UNK 05-NOV-2020
DEFINITION  Pichia ENO 3'-Homology Region in pOpen-v3.
ACCESSION   BBF10K_000460
VERSION     BBF10K_000460.1
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 2645)
  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..2545
                     /label="Pichia ENO 3'-Homology Region 493bp"
                     /original_record="part"
     protein_bind    2546..2553
                     /label="NotI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
                     /original_record="part"
     misc_feature    2002..2021
                     /label="FreeGenes dialout primer"
                     /ApEinfo_fwdcolor="#0048BA"
                     /ApEinfo_revcolor="#0048BA"
     misc_feature    complement(2566..2585)
                     /label="FreeGenes dialout primer"
                     /ApEinfo_fwdcolor="#0048BA"
                     /ApEinfo_revcolor="#0048BA"
     misc_feature    2586..2645
                     /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(2555..2558,2560..2565))
                     /label="BsaI"
                     /ApEinfo_fwdcolor="#00FF2C"
                     /ApEinfo_revcolor="#00FF2C"
     protein_bind    2607..2624
                     /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 gaagaattac tgacccctcg gggtctctag acactcgtat
     2041 cgcatgtcgg tgcaccgtaa atttgccttg caattggccc taaccacgga tggttaactt
     2101 caagccatca cgacagcaat tgagtcggcg catagctacc ctcctcttct tgaccccatg
     2161 cataggacca accttaaccg atggaacagg ttcctccgct ccgtcccctg gtagtgtctc
     2221 tgcgcaagaa atagttaagg tattaacact gatctctcgc acccccctca cagtactgtt
     2281 atggtgaatt gacaaagcca ttggctagat tgaaacatgt aattcatatg taatcttgtt
     2341 caattaacga gcttcgtaca gtctcaatct agacgtctga taatggcgtt tgtgctccta
     2401 atcgatgagc catctcatgt gacgtctata cgcttcgatg gcttccgtcg cgaatataga
     2461 accacttgaa atatgctgca aaccacgatc caccctggtc ctgaaaagat ataaatacag
     2521 cacatctagc aggcttttgc gctgcgcggc cgcacgaatg agaccaagga actatggcat
     2581 cgagcgttat gggcgccgga cgctactagg gataacaggg taatattgcg acacccatct
     2641 ccagc
//
