LOCUS       sequence-133799-        3185 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Level 2 Destination vector - Position A - Assembly of 4-5 TU's from 
            Level 1-ABCD or ABCDE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3185)
  AUTHORS   Moore SJ, Lai HE, Kelwick RJ, Chee SM, Bell DJ, Polizzi KM, Freemont
            PS
  TITLE     EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2016 May 2.
  PUBMED    27096716
REFERENCE   2  (bases 1 to 3185)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2016 May 2."
COMMENT     Sequence Label: pTU2-A-RFP
FEATURES             Location/Qualifiers
     source          1..3185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    135..156
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(171..176,177..194,195..201)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    209..225
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             243..254
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             261..938
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      983..1054
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1070..1097
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1138..1195
                     /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(1219..1236)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1390..1978)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1470..1489)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2161..2255)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2276..2935)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2850..2869
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3097..3115
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3119..3162)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(3165..3185,1)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggtctcacta tatctagaga cgtctagaca tatgcaatac gcaaaccgcc tctccccgcg
       61 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      121 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      181 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacatactag
      241 agaaagagga gaaatactag atggcttcct ccgaagacgt tatcaaagag ttcatgcgtt
      301 tcaaagttcg tatggaaggt tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg
      361 aaggtcgtcc gtacgaaggt acccagaccg ctaaactgaa agttaccaaa ggtggtccgc
      421 tgccgttcgc ttgggacatc ctgtccccgc agttccagta cggttccaaa gcttacgtta
      481 aacacccggc tgacatcccg gactacctga aactgtcctt cccggaaggt ttcaaatggg
      541 aacgtgttat gaacttcgaa gacggtggtg ttgttaccgt tacccaggac tcctccctgc
      601 aagacggtga gttcatctac aaagttaaac tgcgtggtac caacttcccg tccgacggtc
      661 cggttatgca gaaaaaaacc atgggttggg aagcttccac cgaacgtatg tacccggaag
      721 acggtgctct gaaaggtgaa atcaaaatgc gtctgaaact gaaagacggt ggtcactacg
      781 acgctgaagt taaaaccacc tacatggcta aaaaaccggt tcagctgccg ggtgcttaca
      841 aaaccgacat caaactggac atcacctccc acaacgaaga ctacaccatc gttgaacagt
      901 acgaacgtgc tgaaggtcgt cactccaccg gtgcttaata acgctgatag tgctagtgta
      961 gatcgctact agagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1021 tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc
     1081 gggtgggcct ttctgcgttt atagcatgcc gtctcattag gtacagagac cctgcagtcc
     1141 ggcaaaaaag ggcaaggtgt caccaccctg ccctttttct ttaaaaccga aaagattact
     1201 tcgcgttatg caggcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1381 gctggcgttt ttccacaggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1561 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1801 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct cgaggcttgg
     2161 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     2221 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg
     2281 ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga
     2341 agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt
     2401 gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt
     2461 ttaaatcaaa actggtgaaa ctcacccagg gattggctga cacgaaaaac atattctcaa
     2521 taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata
     2581 tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag
     2641 tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt
     2701 ctttcattgc catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa
     2761 aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca
     2821 gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt
     2881 tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag
     2941 cttccttagc tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt
     3001 cattatggtg aaagttggaa cctcttacgt gcccgatcaa ctcgagtgcc acctgacgtc
     3061 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
     3121 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt
     3181 ctaga
//
