LOCUS       sequence-338675-        5401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  StrepFlex MoClo vector for cloning in E. coli and Streptomyces 
            expression - equivalent of pTU1-A-RFP from EcoFlex (ACS Synth. Biol.
            2016, 5, 10, 1059–1069).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5401)
  AUTHORS   Moore SJ, Lai HE, Chee SM, Toh M, Coode S, Chengan K, Capel P, Corre
            C, de Los Santos EL, Freemont PS
  TITLE     A Streptomyces venezuelae Cell-Free Toolkit for Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2021 Feb 19;10(2):402-411. doi: 
            10.1021/acssynbio.0c00581. Epub 2021 Jan 26.
  PUBMED    33497199
REFERENCE   2  (bases 1 to 5401)
  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"; doi: 
            "10.1021/acssynbio.0c00581"; journalName: "ACS Synth Biol"; date: 
            "2021-02-19- 19"; volume: "10"; issue: "2"; pages: "402-411"
COMMENT     Sequence Label: pSF1C-A-RFP
FEATURES             Location/Qualifiers
     source          1..5401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     38..56
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(60..103)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    106..125
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     protein_bind    250..271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(286..291,292..309,310..316)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    324..340
                     /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             358..369
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             376..1053
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      1098..1169
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1185..1212
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1252..1269)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1417..2005)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1497..1516)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2209..3009
                     /codon_start=1
                     /gene="aac(3)IV"
                     /product="aminoglycoside 3-N-acetyltransferase type IV     
                               "
                     /label=ApmR
                     /note="confers resistance to apramycin (Paget and Davies, 
                     1996)"
                     /translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
                     LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
                     KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
                     LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
                     AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
     oriT            3186..3295
                     /note="incP origin of transfer"
ORIGIN
        1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggcagaa
       61 tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt cgcggccgct
      121 tctagaacgt ctcaatctct atagagaccc aatacgcaaa ccgcctctcc ccgcgcgttg
      181 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      241 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
      301 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacat actagagaaa
      361 gaggagaaat actagatggc ttcctccgaa gacgttatca aagagttcat gcgtttcaaa
      421 gttcgtatgg aaggttccgt taacggtcac gagttcgaaa tcgaaggtga aggtgaaggt
      481 cgtccgtacg aaggtaccca gaccgctaaa ctgaaagtta ccaaaggtgg tccgctgccg
      541 ttcgcttggg acatcctgtc cccgcagttc cagtacggtt ccaaagctta cgttaaacac
      601 ccggctgaca tcccggacta cctgaaactg tccttcccgg aaggtttcaa atgggaacgt
      661 gttatgaact tcgaagacgg tggtgttgtt accgttaccc aggactcctc cctgcaagac
      721 ggtgagttca tctacaaagt taaactgcgt ggtaccaact tcccgtccga cggtccggtt
      781 atgcagaaaa aaaccatggg ttgggaagct tccaccgaac gtatgtaccc ggaagacggt
      841 gctctgaaag gtgaaatcaa aatgcgtctg aaactgaaag acggtggtca ctacgacgct
      901 gaagttaaaa ccacctacat ggctaaaaaa ccggttcagc tgccgggtgc ttacaaaacc
      961 gacatcaaac tggacatcac ctcccacaac gaagactaca ccatcgttga acagtacgaa
     1021 cgtgctgaag gtcgtcactc caccggtgct taataacgct gatagtgcta gtgtagatcg
     1081 ctactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
     1141 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     1201 ggcctttctg cgtttatagg tctcatgttt gccagagacg ctgcaggctt cctcgctcac
     1261 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1321 aatttaagcc gaacaggaag cacagctcct actgaacatg tgagcaaaag gccagcaaaa
     1381 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1441 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1501 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1561 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1621 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1681 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1741 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1801 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     1861 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1921 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1981 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2041 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2101 cacctagatc cttttggttc atgtgcagct ccatcagcaa aaggggatga taagtttatc
     2161 accaccgact atttgcaaca gtgccgttga tcgtgctatg atcgactgat gtcatcagcg
     2221 gtggagtgca atgtcgtgca atacgaatgg cgaaaagccg agctcatcgg tcagcttctc
     2281 aaccttgggg ttacccccgg cggtgtgctg ctggtccaca gctccttccg tagcgtccgg
     2341 cccctcgaag atgggccact tggactgatc gaggccctgc gtgctgcgct gggtccggga
     2401 gggacgctcg tcatgccctc gtggtcaggt ctggacgacg agccgttcga tcctgccacg
     2461 tcgcccgtta caccggacct tggagttgtc tctgacacat tctggcgcct gccaaatgta
     2521 aagcgcagcg cccatccatt tgcctttgcg gcagcggggc cacaggcaga gcagatcatc
     2581 tctgatccat tgcccctgcc acctcactcg cctgcaagcc cggtcgcccg tgtccatgaa
     2641 ctcgatgggc aggtacttct cctcggcgtg ggacacgatg ccaacacgac gctgcatctt
     2701 gccgagttga tggcaaaggt tccctatggg gtgccgagac actgcaccat tcttcaggat
     2761 ggcaagttgg tacgcgtcga ttatctcgag aatgaccact gctgtgagcg ctttgccttg
     2821 gcggacaggt ggctcaagga gaagagcctt cagaaggaag gtccagtcgg tcatgccttt
     2881 gctcggttga tccgctcccg cgacattgtg gcgacagccc tgggtcaact gggccgagat
     2941 ccgttgatct tcctgcatcc gccagaggcg ggatgcgaag aatgcgatgc cgctcgccag
     3001 tcgattggct gagctcatga gcggagaacg agatgacgtt ggaggggcaa ggtcgcgctg
     3061 attgctgggg caacacgtgg agcggatcgg ggattgtctt tcttcagctc gctgatgata
     3121 tgccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc
     3181 ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     3241 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     3301 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     3361 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc agggttatgc
     3421 agcggaaaag atccgtcgac ctgcaggcat gctcctctag agtccgtctg acgcgtgtgg
     3481 ggagcacgcc ggcccgctcg gtagacagca ctctacgcat tgtggctgct gtgcacatca
     3541 ctctttcgag cgacgtgatc cgcaggattc cgtggggatg tggcatctat gtggcatccg
     3601 ggtacgacaa cggccccccg ttcgaatgga acggggggcc gttgactccc tgacctgcga
     3661 tgcaggttgt gcccccggca ggattcgaac ctgcgacacc cgctttagga gagaacgtaa
     3721 agcgatcatg agcctcagtg agtctgagtg agtctcagtg agcctgtcgc cataggggct
     3781 tcacctgcat aaatgagttt cagtgagtct cggtgagtct cagtgagggg gtgttgtggc
     3841 atccctgtgg catctgcgtg gcatccggcg tcacaccgcc cgcagcgttc gaacaccctc
     3901 ggtttcggga gccatcgccg cccgcacctt cgccgccacg tcggccggcg aatgctggta
     3961 caaccacgtc accttcgacc ctcgctcgtg acccatcacg gtctgcgtgt ccttctcgtc
     4021 aatgcccaca tccttgagcc acgtggcgaa cacgtggcga aggtcgtgca cccgcggcca
     4081 ccactccatg cggccggtgt cccggttctt cacctcgcgg gcgaggcctg cagccttgat
     4141 tgcagggatc catgtgcgcc ggaagttgtg ccgggtcagg acgccgccct gtgggccgcg
     4201 gaacacgagc tcatccggcg cgaggtcgta cgggtcctcg ccaatcgggg tgatcgtggg
     4261 agtcggccgc caccgggtga ccatggtgcg gatcgcgtcc tcggcctgcg gcgtgagcgg
     4321 gaccgtgcgg aacccggcgt tgctcttcgg cgcaggcttc cggaacagct tgccttcgtc
     4381 gtcactgagc acttccttca ccttgaggtg gtgggcgtcc aggtcgacgt tctcccagcg
     4441 cagcgccgtc gcctcgcccc accggagtcc ggtgtgttca aggaagacga cgagcggccg
     4501 gtagtacatc ggcaggtgac tgacgatcag cgcgcactgc gcgcggtcgg gcggcatcag
     4561 ttcgtccgga tgcttcgccg gcgcctcgcc aatgtccagg tcggccgccg ggttgaacgg
     4621 gatacgccgg ccgtccttga cggcggcccg gagcatcgag ttcagcacct catgaacctt
     4681 cttccgggtg tggtagccct tcacctcctt cgtgatccac gcctgcagct cgatgtactc
     4741 caagtcgcac agacgccact gcccccactt cggctcgatg tgcgcggccc agttcgaccg
     4801 cttccggttg gcggtcgtga ctgctcggtc cggctgcgcc tcccaccaca gtttccacca
     4861 ctgggacagg gtgatctcgc cacgcttcgg gtcggcgtac gtccgttcgc ggacctgcgt
     4921 gcggatcttg tcgaggtgcg cctcggctgc cttcttaccg ccgtccttga tcgcaaagca
     4981 tttctgcttc tggttgccgg ccgggtcgcg gtaacgcgcc tgccacgagc cggtgcagtc
     5041 gcgtcggcgg ttccgctctc cgtacttctc gggcgggtac ttctccatgc acagctggca
     5101 gccgcagctc ttcgccctca actgccgtgg gttgttcgtc gctctaggcg ccatgggtca
     5161 cctgtccgct cctccgctgt tcgggaacgc ggggcaggat gtgtacggct gccccacacc
     5221 agcagactgc accgtccggg ggctgctcta cagccagttc ttcaaggacg gctcgaaggg
     5281 ctacgaggga agcatcccgg gggagggtgg caggcagggt gaccgtgttt ccagcggcgt
     5341 cgtacggccg gatgacactc gttcgttcac acgttgcagc agagtacttg ccacctgacg
     5401 t
//
