LOCUS       sequence-492925-        2355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Full transcription unit stuffer in an alpha-level 
            cloning vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2355)
  AUTHORS   Cooke MB, Welch KT, Deus Ramirez L, Wen AX, Marciano DC,, Herman C
  TITLE     GoldenBraid2.0 E. coli: A comprehensive and characterized toolkit 
            for enterics.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2355)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBSKa2_Stuffer2
FEATURES             Location/Qualifiers
     source          1..2355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(155..743)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(235..254)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        858..960
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             961..1776
                     /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             961..1776
                     /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(1034..1053)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1890..2345
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1977..1996)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2187..2208
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgtctcagtc aggaggatcg ctctgccatc tagcacgctt gagacgaagc ttgacacggt
       61 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      121 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      181 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      241 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      301 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      361 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      421 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      481 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
      541 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
      601 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
      661 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
      721 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
      781 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
      841 cctagatcct tttgtgatga tcggcacgta agaggttcca actttcacca taatgaaata
      901 agatcactac cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa
      961 atgagccata ttcaacggga aacgtcttgc tcaaggccgc gattaaattc caacatggat
     1021 gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc
     1081 taccgattgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc
     1141 gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct
     1201 cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg
     1261 atcccaggga aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt
     1321 gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct
     1381 tttaacagcg atcgcgtatt tcgactcgct caggcgcaat cacgaatgaa taacggtttg
     1441 gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa
     1501 gaaatgcata aacttttgcc attctcaccg gattcagtag tcactcatgg tgatttctca
     1561 cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc
     1621 ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct
     1681 ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa
     1741 ttgcagtttc atttgatgct cgatgagttt ttctgatttt ttttaaggca gttattggtg
     1801 cccttaaacg cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     1861 ttcgatgata agctgtcaaa catgatgtta cgcgccctgt agcggcgcat taagcgcggc
     1921 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1981 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2041 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2101 tgattagggt gatggttcac gtagtgggcc agcgccctga tagacggttt ttcgcccttt
     2161 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2221 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2281 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     2341 aattttaaca agctt
//
