LOCUS       sequence-240452-        2812 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Alpha 2 destination vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2812)
  AUTHORS   Sarrion-Perdigones A, Chang L, Gonzalez Y, Gallego-Flores T, Young 
            DW, Venken KJT
  TITLE     Examining multiple cellular pathways at once using multiplex 
            hextuple luciferase assaying.
  JOURNAL   Nat Commun. 2019 Dec 13;10(1):5710. doi: 10.1038/s41467-019-13651-y.
  PUBMED    31836712
REFERENCE   2  (bases 1 to 2812)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-12-13"; volume: "10(1):5710. doi"; pages: " 
            10.1038/s41467-019-13651-y"
COMMENT     Sequence Label: Alpha2
FEATURES             Location/Qualifiers
     source          1..2812
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        63..81
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     63..80
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    88..144
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(146..165)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(147..165)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(172..189)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(172..188)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(181..203)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(397..416)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(548..1136)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(628..647)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1251..1353
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1354..2169
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1354..2169
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1427..1446)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2283..2738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2370..2389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2580..2601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        join(2774..2812,1..9)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
ORIGIN
        1 taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgaccatga ttacgccaag
       61 ctatttaggt gacactatag aatactcaag cttgcatgcc tgcaggtcga ctctagagga
      121 tccccgggta ccgagctcga attcgcccta tagtgagtcg tattacaatt cactggccgt
      181 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      241 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      301 acagttgcgc agcctgaatg gcgaatgacg cctgatgcgg tattttctcc ttacgcatct
      361 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
      421 gggtctctcg cttgagacga agcttgacac ggttatccac agaatcaggg gataacgcag
      481 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      541 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      601 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      661 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      721 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      781 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      841 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      901 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      961 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1021 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1081 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1141 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1201 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgtga tgatcggcac
     1261 gtaagaggtt ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag
     1321 ttatcgagat tttcaggagc taaggaagct aaaatgagcc atattcaacg ggaaacgtct
     1381 tgctcaaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     1441 cgcgataatg tcgggcaatc aggtgcgaca atctaccgat tgtatgggaa gcccgatgcg
     1501 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     1561 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     1621 actcctgatg atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta
     1681 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     1741 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgactc
     1801 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     1861 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     1921 ccggattcag tagtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     1981 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     2041 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     2101 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     2161 tttttctgat tttttttaag gcagttattg gtgcccttaa acgcctggtt gctacgcctg
     2221 aataagtgat aataagcgga tgaatggcag aaattcgatg ataagctgtc aaacatgatg
     2281 ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2341 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2401 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2461 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2521 gccagcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2581 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2641 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2701 taacgcgaat tttaacaaaa tattaacgct tacaatttta acaagcttcg tctcagtcag
     2761 gagagagacc cctgttgaca attaatcatc ggcatagtat atcggcatag ta
//
