LOCUS       pENG-087_Leu2_p4        6522 bp DNA     circular SYN 26-MAR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6522)
  AUTHORS   Atti English
  TITLE     Direct Submission
  JOURNAL   Exported Dec 12, 2023 from SnapGene 7.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6522
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     protein_bind    1..4
                     /label=BsaI
     misc_feature    5..504
                     /label=LEU2 5' Homology
     misc_feature    complement(505..524)
                     /label=LEU2 5' barcode
     protein_bind    525..528
                     /label=BsaI
     protein_bind    complement(join(544..547,549..554))
                     /label=BsmBI
     misc_feature    555..697
                     /label=ConS
     primer_bind     602..619
                     /label=oENG525_seq
     primer_bind     698..721
                     /label=oENG314_ytk
     protein_bind    698..701
                     /label=BsaI
     misc_feature    702..707
                     /label=spacer1
     promoter        708..721
                     /label=DBM_42-10_repeat_unit
                     /note="repeat this in the 5' direction to add additional 
                     43-8 binding sites
                     "
     misc_feature    708..718
                     /label=DBM_sTF_42-10
     misc_feature    719..721
                     /label=spacer2_recode
     promoter        722..735
                     /label=DBM_42-10_repeat_unit
                     /note="repeat this in the 5' direction to add additional 
                     43-8 binding sites
                     "
     misc_feature    722..732
                     /label=DBM_sTF_42-10
     misc_feature    733..735
                     /label=spacer2_recode
     promoter        736..749
                     /label=DBM_42-10_repeat_unit
                     /note="repeat this in the 5' direction to add additional 
                     43-8 binding sites
                     "
     misc_feature    736..746
                     /label=DBM_sTF_42-10
     misc_feature    747..749
                     /label=spacer2
     misc_feature    750..760
                     /label=DBM_sTF_42-10
     misc_feature    761..766
                     /label=spacer3
     promoter        767..1003
                     /label=miniCyc1_promoter
     primer_bind     complement(982..1020)
                     /label=oENG315_ytk
     primer_bind     complement(986..1016)
                     /label=oENG109
     misc_feature    1004..1009
                     /label=spacer4
     5'UTR           1010..1016
                     /label=kozak+start_codon
     primer_bind     1016..1038
                     /label=oENG518_ytk
     protein_bind    1017..1020
                     /label=_3
                     /label=BsaI
                     /bound_moiety="BsaI"
     CDS             1018..1725
                     /codon_start=1
                     /product="Bright green-yellow fluorescent protein derived 
                     from Branchiostoma lanceolatum"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     primer_bind     1018..1045
                     /label=MA_065_gib
     primer_bind     1018..1038
                     /label=MA_o80_PKP40_seq
     primer_bind     complement(1062..1084)
                     /label=oENG524_seq
     primer_bind     1314..1333
                     /label=oENG520_seq
     primer_bind     complement(1699..1725)
                     /label=MA_o66_gib
     primer_bind     complement(1705..1732)
                     /label=oENG519_ytk
     primer_bind     complement(1705..1728)
                     /label=mNG_pGKL_R
     primer_bind     complement(1705..1725)
                     /label=MA_o87
     protein_bind    1728..1731
                     /label=BsaI
     terminator      1741..1964
                     /label=ScTDH1 Terminator
     primer_bind     complement(1909..1931)
                     /label=oENG523_seq
     protein_bind    1965..1968
                     /label=BsaI
     misc_feature    1969..2111
                     /label=ConE
     primer_bind     complement(2038..2058)
                     /label=oENG526_seq
     protein_bind    join(2112..2117,2119..2122)
                     /label=BsmBI
     misc_feature    2119..2143
                     /label=ConE scar
     protein_bind    2159..2162
                     /label=BsaI
     promoter        2163..2810
                     /label=LEU2 Promoter
     primer_bind     complement(2770..2813)
                     /label=org349_leu2_tag_2
     CDS             2811..3905
                     /label=LEU2
     primer_bind     2814..2834
                     /label=org348_leu2_tag_1
     primer_bind     2814..2834
                     /label=org370_leu2_ttrz_f
     primer_bind     complement(3877..3905)
                     /label=org366_leu2_r_bamhi
     terminator      3906..4079
                     /label=AgLEU2 Terminator
     protein_bind    4080..4083
                     /label=BsaI
     misc_feature    4084..4103
                     /label=LEU2 3' barcode
     misc_feature    4104..4593
                     /label=LEU2 3' Homology
     protein_bind    4594..4597
                     /label=BsaI
     CDS             complement(4606..5616)
                     /label=SpecR
     promoter        complement(5617..5750)
                     /label=SpecR Promoter
     rep_origin      complement(5751..6514)
                     /direction=LEFT
                     /label=ColE1
ORIGIN
        1 caatacccag aagaacagta aaataaagca aggtacgtga aattaatatt tttaaatggt
       61 tctaaccgat gccgaagaac tgcgcagtcc ggttataacg tctgacatgt ccttttttga
      121 tttggaatcc aaccactcaa gtgactctgt tcatttactt tgcgaaaaat atacccacaa
      181 attgcccatc gaaagtgaat cgcaaaccac cttcagactg gcaccgacaa agcaaagatt
      241 atacagacag agtactttat acgtaccgtt aagtctcaag caaagggttt tcttatttac
      301 tgaacgggta aagagtatct gggccggctt gccaagatgc aaaccgaata agtatttcaa
      361 agttgcattt gccttagccg tcctgacacc attggctatt tggatatttt atattgactt
      421 tcgtgtacat tgatcacatc gactgttcta ttggcaaatg aaccacgggc attgactatt
      481 tttcaggtta ctactatata ttatcatcac gggcaaggat tgtaccctga attcgcatct
      541 agactgatga gacgtggtag agccacaaac agccggtaca agcaacgatc tccaggacca
      601 tctgaatcat gcgcggatga cacgaactca cgacggcgat cacagacatt aacccacagt
      661 acagacactg cgacaacgtg gcaattcgtc gcaatacaac ggaattcaga cgctgctctt
      721 cagacgctgc tctgcagacg ctgctctcca gacgctgctc ggatcccaga tccgccaggc
      781 gtgtatatat agcgtggatg gccaggcaac tttagtgctg acacatacag gcatatatat
      841 atgtgtgcga cgacacatga tcatatggca tgcatgtgct ctgtatgtat ataaaactct
      901 tgttttcttc ttttctctaa atattctttc cttatacatt aggacctttg cagcataaat
      961 tactatactt ctatagacac acaaacacaa atacacacac taatctagat attaaatatg
     1021 gtttcaaagg gtgaagagga taacatggca agtttgccgg ccacacatga acttcacatt
     1081 ttcgggagca taaatggcgt tgattttgac atggtcggtc aggggactgg gaaccccaac
     1141 gacggatacg aagaactcaa cttgaaatct accaaaggtg atctccaatt ttcaccgtgg
     1201 attcttgttc cgcacattgg gtatggattt catcagtatt tgccataccc tgatggcatg
     1261 tcacccttcc aggccgctat ggtcgatggt agcggctacc aggtacaccg cacaatgcag
     1321 tttgaagacg gtgcatcact gaccgtgaac taccgatata cctatgaggg ttcccatatt
     1381 aaaggagaag ctcaagtaaa gggaacggga ttcccggccg atggtcccgt gatgactaac
     1441 tccttgacag cagcagactg gtgtcgctct aagaagactt atcccaacga taaaaccatt
     1501 ataagtacat ttaaatggag ttatacaacg ggcaacggta agaggtacag aagcacggca
     1561 cggaccactt acaccttcgc taagccgatg gccgcaaact acttgaagaa ccaaccgatg
     1621 tacgtattcc gaaagacgga actgaaacat agcaaaactg aactgaattt caaggagtgg
     1681 cagaaagcgt ttactgatgt catgggtatg gacgagctgt ataagggatc ctaactcgag
     1741 ataaagcaat cttgatgagg ataatgattt ttttttgaat atacataaat actaccgttt
     1801 ttctgctaga ttttgtgatg acgtaaataa gtacatatta ctttttaagc caagacaaga
     1861 ttaagcatta actttaccct tttctttcta agtttcaata ttagttatca ctgtttaaaa
     1921 gttatggcga gaacgtcggc ggttaaaata tattaccctg aacggctgga aatctgctcg
     1981 tcagtggtgc tcacactgac gaatcatgta cagatcatac cgatgactgc ctggcgactc
     2041 acaactaagc aagacagccg gaaccagcgc cggcgaacac cactgcatat atggcatatc
     2101 acaacagtcc acgtctcaag cagttacaga gatgttacga accactagtg cactgcagta
     2161 catcgaggag aacttctagt atatctacat acctaatatt attgccttat taaaaatgga
     2221 atcccaacaa ttacatcaaa atccacattc tcttcaaaat caattgtcct gtacttcctt
     2281 gttcatgtgt gttcaaaaac gttatattta taggataatt atactctatt tctcaacaag
     2341 taattggttg tttggccgag cggtctaagg cgcctgattc aagaaatatc ttgaccgcag
     2401 ttaactgtgg gaatactcag gtatcgtaag atgcaagagt tcgaatctct tagcaaccat
     2461 tatttttttc ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg
     2521 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg
     2581 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga
     2641 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg
     2701 acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct
     2761 tttacatttc agcaatatat atatatatat ttcaaggata taccattcta atgtctgccc
     2821 ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca gccgaagcca
     2881 ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat ttcgaaaatc
     2941 atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat gaggcgctgg
     3001 aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct aaatggggaa
     3061 ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt caattgtacg
     3121 ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct ccaatcaagc
     3181 cacaatttgc taaaggtact gacttcgttg ttgtcagaga attagtggga ggtatttact
     3241 ttggtaagag aaaggaggac gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg
     3301 ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa catgagccac
     3361 cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga ttatggagaa
     3421 aaactgtgga ggaaaccatc aagaacgaat ttcctacatt gaaggttcaa catcaattga
     3481 ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt attataatca
     3541 ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca ggttccttgg
     3601 gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca tttggtttgt
     3661 acgaaccatg ccacggttct gctccagatt tgccaaagaa taaggtcaac cctatcgcca
     3721 ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg
     3781 ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt gatttaggtg
     3841 gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag aaaatccttg
     3901 cttaacctgg aggacccttc tctttagact attctactct tatgcacgta aaaaattcta
     3961 ggaaatatgt attaactagg agtaaaataa ccggctagtg gcattcatat agccgtctgt
     4021 ttacatctac atcacacatt tcgagtgtat atctcgcaac gttggcgtta aataggcagg
     4081 agtactcgta tcgcatgtcg gtgcgacacg aaattacaaa atggaatatg ttcatagggt
     4141 agacgaaact atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta
     4201 aaggaataca ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc
     4261 actttaacat taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa
     4321 aatcatgaaa ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa
     4381 tacaacaaat aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt
     4441 cttgaaccat ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg
     4501 ccttaacgac gtagtcgacc tcctcttcag tactaaatct accaatacca aatctgatgg
     4561 aagaatgggc taatgcatca tccttaccca gcgccgagcg gccgcttatt tgccgactac
     4621 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
     4681 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
     4741 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
     4801 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
     4861 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
     4921 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
     4981 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
     5041 cgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
     5101 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
     5161 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
     5221 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
     5281 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
     5341 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcataat
     5401 gtttaacttt gttttagggc gactgccctg ctgcgtaaca tcgttgctgc tccataacat
     5461 caaacatcga cccacggcgt aacgcgcttg ctgcttggat gcccgaggca tagactgtac
     5521 cccaaaaaaa cagtcataac aagccatgaa aaccgccact gcgccgttac caccgctgcg
     5581 ttcggtcaag gttctggacc agttgcgtga gcgcatacgc tacttgcatt acagcttacg
     5641 aaccgaacag gcttatgtcc actgggttcg tgccttcatc cgtttccacg gtgtgcgtca
     5701 cccggcaacc ttgggtagca gcgaagtcga ggcatttctg tcctggctgg tcatgaccaa
     5761 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5821 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5881 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5941 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6001 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6061 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6121 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6181 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6241 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6301 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6361 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6421 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6481 tcctgcgtta tcccctgatt ctgtggataa ccgtgcggcc gc
//
