LOCUS       sequence-373958-        5205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of the rep protein of phage MS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5205)
  AUTHORS   Wandera KG, Alkhnbashi OS, Bassett HVI, Mitrofanov A, Hauns S, Migur
            A, Backofen R, Beisel CL
  TITLE     Anti-CRISPR prediction using deep learning reveals an inhibitor of 
            Cas13b nucleases.
  JOURNAL   Mol Cell. 2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003.
  PUBMED    35649413
REFERENCE   2  (bases 1 to 5205)
  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: "Mol Cell. 
            2022 May 24. pii: S1097-2765(22)00437-3. doi: 
            10.1016/j.molcel.2022.05.003."
COMMENT     Sequence Label: pPJ23116-MS2-rep
FEATURES             Location/Qualifiers
     source          1..5205
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     148..166
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        190..224
                     /label=BBa_J23116
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.16"
     terminator      1919..2005
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2543..3493)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3541..3763
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4255..4349)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4380..5174)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4491..4510)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5101..5120
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gatcttgatc ccctgcgcca tcagatcctt ggcggcaaga aagccatcca gtttactttg
       61 cagggcttcc caaccttacc agagggcgcc ccagctggca attccgacgt ctaagaaacc
      121 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
      181 ctcgagttct tgacagctag ctcagtccta gggactatgc tagcggatcc tctagataga
      241 cgccggccat tcaaacatga ggattaccca tgtcgaagac aacaaagaag ttcaactctt
      301 tatgtattga tcttcctcgc gatctttctc tcgaaattta ccaatcaatt gcttctgtcg
      361 ctactggaag cggtgatccg cacagtgacg actttacagc aattgcttac ttaagggacg
      421 aattgctcac aaagcatccg accttaggtt ctggtaatga cgaggcgacc cgtcgtacct
      481 tagctatcgc taagctacgg gaggcgaatg gtgatcgcgg tcagataaat agagaaggtt
      541 tcttacatga caaatccttg tcatgggatc cggatgtttt acaaaccagc atccgtagcc
      601 ttattggcaa cctcctctct ggctaccgat cgtcgttgtt tgggcaatgc acgttctcca
      661 acggtgctcc tatggggcac aagttgcagg atgcagcgcc ttacaagaag ttcgctgaac
      721 aagcaaccgt taccccccgc gctctgagag cggctctatt ggtccgagac caatgtgcgc
      781 cgtggatcag acacgcggtc cgctataacg agtcatatga atttaggctc gttgtaggga
      841 acggagtgtt tacagttccg aagaataata aaatagatcg ggctgcctgt aaggagcctg
      901 atatgaatat gtacctccag aaaggggtcg gtgctttcat cagacgccgg ctcaaatccg
      961 ttggtataga cctgaatgat caatcgatca accagcgtct ggctcagcag ggcagcgtag
     1021 atggttcgct tgcgacgata gacttatcgt ctgcatccga ttccatctcc gatcgcctgg
     1081 tgtggagttt tctcccacca gagctatatt catatctcga tcgtatccgc tcacactacg
     1141 gaatcgtaga tggcgagacg atacgatggg aactattttc cacaatggga aatgggttca
     1201 catttgagct agagtccatg atattctggg caatagtcaa agcgacccaa atccattttg
     1261 gtaacgccgg aaccataggc atctacgggg acgatattat atgtcccagt gagattgcac
     1321 cccgtgtgct agaggcactt gcctactacg gttttaaacc gaatcttcgt aaaacgttcg
     1381 tgtccgggct ctttcgcgag agctgcggcg cgcactttta ccgtggtgtc gatgtcaaac
     1441 cgttttacat caagaaacct gttgacaatc tcttcgccct gatgctgata ttaaatcggc
     1501 tacggggttg gggagttgtc ggaggtatgt cagatccacg cctctataag gtgtgggtac
     1561 ggctctcctc ccaggtgcct tcgatgttct tcggtgggac ggacctcgct gccgactact
     1621 acgtagtcag cccgcctacg gcagtctcgg tatacaccaa gactccgtac gggcggctgc
     1681 tcgcggatac ccgtacctcg ggtttccgtc ttgctcgtat cgctcgagaa cgcaagttct
     1741 tcagcgaaaa gcacgacagt ggtcgctaca tagcgtggtt ccatactgga ggtgaaatca
     1801 ccgacagcat gaagtccgcc ggcgtgcgcg ttatacgcac ttcggagtgg ctaacgccgg
     1861 ttcccacatt ccctcaggag tgtgggccag cgagctctcc tcggtagtct agaggcatca
     1921 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     1981 gaacgctctc ctgagtagga caaatccgcc gccctagacc tagggtacgg gttttgctgc
     2041 ccgcaaacgg gctgttctgg tgttgctagt ttgttatcag aatcgcagat ccggcttcag
     2101 ccggtttgcc ggctgaaagc gctatttctt ccagaattgc catgattttt tccccacggg
     2161 aggcgtcact ggctcccgtg ttgtcggcag ctttgattcg ataagcagca tcgcctgttt
     2221 caggctgtct atgtgtgact gttgagctgt aacaagttgt ctcaggtgtt caatttcatg
     2281 ttctagttgc tttgttttac tggtttcacc tgttctatta ggtgttacat gctgttcatc
     2341 tgttacattg tcgatctgtt catggtgaac agctttgaat gcaccaaaaa ctcgtaaaag
     2401 ctctgatgta tctatctttt ttacaccgtt ttcatctgtg catatggaca gttttccctt
     2461 tgatatgtaa cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat
     2521 agatacaaga gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg
     2581 tggttcgttg tttttgcgtg agccatgaga acgaaccatt gagatcatac ttactttgca
     2641 tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt
     2701 gtagtgtttt tcttagtccg ttatgtaggt aggaatctga tgtaatggtt gttggtattt
     2761 tgtcaccatt catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat
     2821 ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt
     2881 tcatattgct gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc
     2941 ttttaaactc atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa
     3001 tctctatatt tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa
     3061 tcctcataga gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt
     3121 tttttaactg gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc
     3181 ttgagaactt ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc
     3241 tgatttccac agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat
     3301 tttccctact gatgttcatc atctgagcgt attggttata agtgaacgat accgtccgtt
     3361 ctttccttgt agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct
     3421 tggtttcatg ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa
     3481 ctaattcaga catacatctc aattggtcta ggtgatttta atcactatac caattgagat
     3541 gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct
     3601 gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac
     3661 ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat
     3721 aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc
     3781 cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa
     3841 accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg
     3901 tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc
     3961 acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca
     4021 aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg
     4081 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt
     4141 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag
     4201 taaggcagcg gtatcatcaa caggcttacc cgtcttactg tccctagtgc ttggattctc
     4261 accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga
     4321 ggtcattact ggatctatca acaggagtcc aagcgagctc tcgaacccca gagtcccgct
     4381 cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg agcggcgata
     4441 ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa gctcttcagc aatatcacgg
     4501 gtagccaacg ctatgtcctg atagcggtcc gccacaccca gccggccaca gtcgatgaat
     4561 ccagaaaagc ggccattttc caccatgata ttcggcaagc aggcatcgcc atgggtcacg
     4621 acgagatcct cgccgtcggg catgcgcgcc ttgagcctgg cgaacagttc ggctggcgcg
     4681 agcccctgat gctcttcgtc cagatcatcc tgatcgacaa gaccggcttc catccgagta
     4741 cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg ggcaggtagc cggatcaagc
     4801 gtatgcagcc gccgcattgc atcagccatg atggatactt tctcggcagg agcaaggtga
     4861 gatgacagga gatcctgccc cggcacttcg cccaatagca gccagtccct tcccgcttca
     4921 gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg tggccagcca cgatagccgc
     4981 gctgcctcgt cctgcagttc attcagggca ccggacaggt cggtcttgac aaaaagaacc
     5041 gggcgcccct gcgctgacag ccggaacacg gcggcatcag agcagccgat tgtctgttgt
     5101 gcccagtcat agccgaatag cctctccacc caagcggccg gagaacctgc gtgcaatcca
     5161 tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt gatca
//
