LOCUS       pSpy0C2                 5618 bp DNA     circular SYN 20-MAR-2024
DEFINITION  Golden Gate ligation product.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5618)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 20, 2024 from SnapGene Viewer 6.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5618
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     source          1373..2371
                     /focus
                     /strain="SF370"
                     /serotype="M1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160490"
                     /organism="Streptococcus pyogenes M1 GAS"
     source          1373..1376
                     /focus
                     /strain="SF370"
                     /serotype="M1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160490"
                     /organism="Streptococcus pyogenes M1 GAS"
     source          2368..2371
                     /focus
                     /strain="SF370"
                     /serotype="M1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160490"
                     /organism="Streptococcus pyogenes M1 GAS"
     source          join(2914..3395,2372..2913)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3396..4404
                     /focus
                     /strain="SF370"
                     /serotype="M1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160490"
                     /organism="Streptococcus pyogenes M1 GAS"
     source          3396..3399
                     /focus
                     /strain="SF370"
                     /serotype="M1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160490"
                     /organism="Streptococcus pyogenes M1 GAS"
     source          4401..4404
                     /focus
                     /strain="SF370"
                     /serotype="M1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:160490"
                     /organism="Streptococcus pyogenes M1 GAS"
     source          4421..4455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          4551..5585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     -35_signal      483..488
                     /standard_name="-35"
                     /label=-35
     -10_signal      506..511
                     /standard_name="-10"
                     /label=-10
     CDS             625..1275
                     /standard_name="Cm(R)"
                     /label=Cm(R)
     terminator      complement(1324..1357)
                     /standard_name="term"
                     /label=term terminator
     unsure          1373..2371
                     /label=Downstream homologous region
                     /note="/modified_by=g_geneious"
                     /note="/standard_name=AmyA FR2 (down) (fragment 2) 
                     (concatenatedsequence 2)"
     rep_origin      2542..3130
                     /label=pBR322 origin
                     /note="/modified_by=g_geneious"
     unsure          3396..4404
                     /label=Upstream homologous region
                     /note="/modified_by=g_geneious"
                     /note="/standard_name=AmyA FR1(up) (fragment 4)
                     (concatenated
                     sequence 4)"
     terminator      4421..4455
                     /standard_name="B - lambda t0 terminator"
                     /label=B - lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    4617..4638
                     /standard_name="CAP binding site"
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the
                     presenceof cAMP."
     promoter        4653..4683
                     /standard_name="lac promoter"
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4691..4707
                     /standard_name="lac operator"
                     /label=lac operator binding site
                     /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             4725..4736
                     /standard_name="RBS"
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4743..5420
                     /codon_start=1
                     /standard_name="mRFP1"
                     /product="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      5465..5536
                     /standard_name="rrnB T1 terminator"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      5552..5579
                     /standard_name="T7Te terminator"
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 atgtgtcata gctgtttcct gcaaagaatg gtgatgacaa ttgtacaggt tatcaataaa
       61 gccattgata ctctcattct tatcccgtat ttattaactt ctcctagtat tagtaattat
      121 catggctgtc atggcgcatt aacggaataa agggtgtgct taaatcgggc cattttgcgt
      181 aataagaaaa aggattaatt atgagcgaat tgaattaata ataaggtaat agatttacat
      241 tagaaaatga aaggggattt tatgcgtgag aatgttacag tctatcccgg caatagttac
      301 ccttattatc aagataagaa agaaaaggat ttttcgctac gctcaaatcc tttaaaaaaa
      361 cacaaaagac cacatttttt aatgtggtct tttattcttc aactaaagca cccattagtt
      421 caacaaacga aaattggata aagtgggata tttttaaaat atatatttat gttacagtaa
      481 tattgacttt taaaaaagga ttgattataa tgaagaaagc agacaagtaa gcctcctaaa
      541 ttcactttag ataaaaattt aggaggcata tcaaatgaac tttaataaaa ttcactttag
      601 ataaaaattt aggaggcata tcaaatgaac tttaataaaa ttgatttaga caattggaag
      661 agaaaagaga tatttaatca ttatttgaac caacaaacga cttttagtat aaccacagaa
      721 attgatatta gtgttttata ccgaaacata aaacaagaag gatataaatt ttaccctgca
      781 tttattttct tagtgacaag ggtgataaac tcaaatacag cttttagaac tggttacaat
      841 agcgacggag agttaggtta ttgggataag ttagagccac tttatacaat ttttgatggt
      901 gtatctaaaa cattctctgg tatttggact cctgtaaaga atgacttcaa agagttttat
      961 gatttatacc tttctgatgt agagaaatat aatggttcgg ggaaattgtt tcccaaaaca
     1021 cctatacctg aaaatgcttt ttctctttct attattccat ggacttcatt tactgggttt
     1081 aacttaaata tcaataataa tagtaattac cttctaccca ttattacagc aggaaaattc
     1141 attaataaag gtaattcaat atatttaccg ctatctttac aggtacatca ttctgtttgt
     1201 gatggttatc atgcaggatt gtttatgaac tctattcagg aattgtcaga taggcctaat
     1261 gactggcttt tataatatga gataatgccg actgtacttt ttacagtcgg ttttctaatg
     1321 tcactaacct gccccgttag ttgaagaagg tttttatatt acagctccag atatgtatgg
     1381 tagagaacca aagacaacag aagcctaata aagccatgtg gctatctgtt atcccaggtt
     1441 tgggacaact atacaataaa caaatcgtta aagggggagt cttattagtt gttttcttcc
     1501 tagagctact agaaattgta gttctaggaa ttcctgcttt aacaggtctt tacagtcttg
     1561 gaagtgttcc aatgcaagat cactccttat ttatgttaat taagggggcc atgcaattaa
     1621 ttactctagt cttatttggt atttttcatt tggttgctat gagagatgct aaattagttg
     1681 cacatcaaat gaatgaaggc aagaaggttc cggtgacggc taaagagacc cttgaagcga
     1741 tttatgaaaa aggatttcct tacttgctca tcattccggc ttatttagca atggctttcg
     1801 ctattatttt tccagtttta gtgaccttgc tgattgcttt taccaattac gattttaggc
     1861 atattcctcc gtatcgttta ttggattggg tagggttgaa gaattttctt aatatcttag
     1921 aattaagcac cttcaaaaca gcatttacat ctgttttgtc atggacgctg atctggacat
     1981 tagcagcgac aagtcttcaa attgttattg gaattgcaac ggctgttatt gctaatcaat
     2041 cctttatcag aggaaaacgt ctctttggtg tgatattctt gttaccatgg gctgttccag
     2101 catttatctc tattatgagt tttggtaatt tctttaatga ttccattggg gccatgaatg
     2161 ttcaggttct tcctttcatc gagaaattcc ttcctttcgt tgacttcgga attgtgtcct
     2221 ggaagacaga ccccttctgg acaaaagtgg ctgttattat ggtgcaaggt tggctaggat
     2281 ttccctatat ctatattttg gtatcaggta ttttacaggc aattccagct gatttatacg
     2341 aagcagcaac cgttgatggt gcgacagcta actgtcagac caagtttact catatatact
     2401 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2461 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2521 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2581 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2641 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2701 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2761 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2821 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2881 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2941 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3001 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3061 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3121 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3181 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3241 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3301 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     3361 atgcagctgg cacgacaggt ttcccgactg gaaagacctg aatcaagatt attctagacg
     3421 ctgtctttaa ccatgccagc agtgatgccg tggaatttca ggatgtcttg agatacggta
     3481 aagagtctaa gttttttgat tggtttatga ctcatgatga acacccatcc atggacctag
     3541 taaactatga aacctttgct ggatgtaact atatgcccaa atggaatacc tcaaatagag
     3601 acgtgcaaga ctatctgatt gagattgggc gatactggat taaagaattt tgtatcgatg
     3661 gttggcgcct agatgtttct gatgaagtat ctcatgattt ttggcgacgt ttccgtcagg
     3721 cggttaaagc ggaaaaagca gatgccattt taattgggga gaactggcat gatgcctatc
     3781 cttatttggc aggggatcag tatgatggga taatgaatta tgctttcacc aaagcctgtt
     3841 tggattactt tgcttttgaa accatgacat caaaacaatt ggctgagaaa ttaagcaaca
     3901 ttttgatgcg aaacacttgg caagtcaatc gcatgaactt gaatcttttg gattctcatg
     3961 atacgcatcg tttcttcact caagttaagg gttccaaaga taaattgtta gctgctttag
     4021 cattgctctt tatctttatg ggcattcctt gcttgtatta tgggacagaa gtggcaatgg
     4081 aagggggtta cgaccctgat tcgcgtcgcg gttttccttg ggatgatgct gagcgggaca
     4141 aggatttctt tagcaaagta aaagcgatta ttgctcttcg ccaacagcca gctcttcaag
     4201 aaggagctat tagcattgat tatgacgaag actgcctcat tctgaaacgc agcttaggtg
     4261 agcaaagcat ttcacttgtt cttaacaatg gtgagagtgc ttattctatt aaggaaggag
     4321 gacagatttt gacaagcaat ggggtaacta aaaacaaaaa acaattactc caggatggat
     4381 atattgtgtt tagtaactaa cagcgaacga atattggata ataaaaaacg cccggcggca
     4441 accgagcgtt ctgaattatg gggatgatgt taaggctatt ggtgtttatg gtaaaacgac
     4501 ggccagtcag cgagaccagc tgaattcaaa aaaaaatcta gatggccggc gattcattaa
     4561 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     4621 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     4681 ttgtgtggaa ttgtgagcgg ataacaattt cacacatact agagaaagag gagaaatact
     4741 agatggcttc ctccgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag
     4801 gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag
     4861 gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca
     4921 tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc
     4981 cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg
     5041 aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct
     5101 acaaagttaa actgcgtggt accaacttcc cgtccgatgg tccggttatg cagaaaaaaa
     5161 ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg
     5221 aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca
     5281 cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg
     5341 acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc
     5401 gtcactccac cggtgcttaa taacgctgat agtgctagtg tagatcgcta ctagagccag
     5461 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     5521 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     5581 ttataaccgg ttaatactag tactgcagtg aaggtctc
//
