LOCUS       sequence-487823-        2367 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Ribosome binding site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2367)
  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  JOURNAL   Unpublished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 2367)
  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            "Unpublished"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: pUPD3_RBS_RBS4
FEATURES             Location/Qualifiers
     source          1..2367
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             18..26
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(72..90)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(73..90)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(106..122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(106..122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        940..1042
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1043..1702
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1109..1128)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1815..2270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1902..1921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2112..2133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2289..2306
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2290..2306
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2313..2332
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2313..2331
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcgcaa agaaaagtaa ggaggtgcgg caatgcgaga ccactcatcg cccaatcact
       61 agtgaattcg ttctatagtg tcacctaaat accatagagc cgttggtcat agctgtttcc
      121 tgtccaggca tcatgacacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttgtgat gatcggcacg taagaggttc
      961 caactttcac cataatgaaa taagatcact accgggcgta ttttttgagt tatcgagatt
     1021 ttcaggagct aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat
     1081 atcccaatgg catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta
     1141 taaccagacc gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca
     1201 caagttttat ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt
     1261 ccgtatggca atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac
     1321 cgttttccat gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt
     1381 ccggcagttt ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta
     1441 tttccctaaa gggtttattg agaatatgtt tttcgtgtca gccaatccct gggtgagttt
     1501 caccagtttt gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat
     1561 gggcaaatat tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca
     1621 tgccgtttgt gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga
     1681 tgagtggcag ggcggggcgt aattttttta aggcagttat tggtgccctt aaacgcctgg
     1741 ttgctacgcc tgaataagtg ataataagcg gatgaatggc agaaattcga tgataagctg
     1801 tcaaacatga tgttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     1861 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     1921 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     1981 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     2041 ttcacgtagt gggccagcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     2101 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     2161 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     2221 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt taacattgta
     2281 gaaacgcatg taaaacgacg gccagtgaat tgtaatacga ctcactatag ggtgaattcc
     2341 gcttaggacc ttagatattg atctcaa
//
