LOCUS       sequence-223027-        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        36..83
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        137..155
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     137..154
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    162..218
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(220..239)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(221..239)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(246..263)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(246..262)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(255..277)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(471..490)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(622..1210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(702..721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1325..1427
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1428..2243
                     /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             1428..2243
                     /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(1501..1520)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2357..2785
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2444..2463)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2654..2675
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 taacaagctt cgtctcagtc aggagagaga cccctgttga caattaatca tcggcatagt
       61 atatcggcat agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgacc
      121 atgattacgc caagctattt aggtgacact atagaatact caagcttgca tgcctgcagg
      181 tcgactctag aggatccccg ggtaccgagc tcgaattcgc cctatagtga gtcgtattac
      241 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
      301 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
      361 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gacgcctgat gcggtatttt
      421 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
      481 tctgatgccg catagggtct ctcgcttgag acgaagcttg acacggttat ccacagaatc
      541 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      601 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      661 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      721 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      781 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      841 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      901 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
      961 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1021 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1081 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1141 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1201 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1261 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1321 gtgatgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     1381 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc
     1441 aacgggaaac gtcttgctca aggccgcgat taaattccaa catggatgct gatttatatg
     1501 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctac cgattgtatg
     1561 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1621 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1681 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa
     1741 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1801 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     1861 gcgtatttcg actcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     1921 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     1981 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2041 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     2101 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     2161 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     2221 tgatgctcga tgagtttttc tgattttttt taaggcagtt attggtgccc ttaaacgcct
     2281 ggttgctacg cctgaataag tgataataag cggatgaatg gcagaaattc gatgataagc
     2341 tgtcaaacat gatgttacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2401 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2461 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2521 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2581 ggttcacgta gtgggccagc gccctgatag acggtttttc gccctttgac gttggagtcc
     2641 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2701 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2761 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat tt
//
