LOCUS       sequence-475781-        2960 bp    DNA     circular SYN 21-JUL-2025
DEFINITION  Cloning gene libraries from oligonucleotide pools.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2960)
  TITLE     Cutler lab 2025
REFERENCE   2  (bases 1 to 2960)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 21, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pPOP-BbsI
FEATURES             Location/Qualifiers
     source          1..2960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     76..94
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(98..141)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    144..165
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     promoter        186..220
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             229..240
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      920..991
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1007..1034
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1077..1134
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1158..1175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1329..1917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1409..1428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2100..2194)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2215..2874)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2789..2808
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 ctcttacgtg cccgatcaac tcgagtgcca cctgacgtct aagaaaccat tattatcatg
       61 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
      121 gctttcgcta aggatgattt ctggaattcg cggccgcttc tagagcgatg agtaatgaag
      181 tcttctttac ggctagctca gtcctaggta tagtgctagc tactagagaa agaggagaaa
      241 tactaaatga gtgtgatcgc taaacaaatg acctacaagg tttatatgtc aggcacggtc
      301 aatggacact actttgaggt cgaaggcgat ggaaaaggta agccctacga gggggagcag
      361 acggtaaagc tcactgtcac caagggcgga cctctgccat ttgcttggga tattttatca
      421 ccacagtgtc agtacggaag cataccattc accaagtacc ctgaggacat ccctgactat
      481 gtaaagcagt cattcccgga gggctataca tgggagagga tcatgaactt tgaagatggt
      541 gcagtgtgta ctgtcagcaa tgattccagc atccaaggca actgtttcat ctaccatgtc
      601 aagttctctg gtttgaactt tcctcccaat ggacctgtca tgcagaagaa aacacagggc
      661 tgggaaccca acactgagcg tctgtttgca cgagatggaa tgctgctagg aaacaacttt
      721 atggctctga agttagaagg aggcggtcac tatttgtgtg aatttaaaac tacttacaag
      781 gcaaagaagc ctgtgaagat gccagggtat cactatgttg accgcaaact ggatgtaacc
      841 aatcacaaca aggattacac ttcggttgag cagtgtgaaa tttccattgc acgcaaacct
      901 gtggtcgcct aataagcttc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
      961 ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg
     1021 tgggcctttc tgcggaagac ttgcttactc atcgcgacta gtagcggccg ctgcagtccg
     1081 gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa aagattactt
     1141 cgcgttatgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1201 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1261 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1321 ctggcgtttt tccacaggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1381 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1441 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1501 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1561 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1621 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1681 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1741 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     1801 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1861 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1921 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1981 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2041 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagctc gaggcttgga
     2101 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
     2161 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
     2221 cccgccctgc cactcgtcgc agtactgttg taattcatta agcattctgc cgacatggaa
     2281 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
     2341 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
     2401 taaatcaaaa ctggtgaaac tcacccaggg attggctgat acgaaaaaca tattctcaat
     2461 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
     2521 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agtgatgaaa acgtttcagt
     2581 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
     2641 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     2701 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     2761 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     2821 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     2881 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     2941 attatggtga aagttggaac
//
