LOCUS       pADH100-Cau             5047 bp    DNA     circular SYN 19-MAR-2025
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5047)
  AUTHORS   Dimitrios
  TITLE     Direct Submission
  JOURNAL   Exported Jul 30, 2025 from SnapGene 8.1.1
            https://www.snapgene.com
COMMENT     Sequence Label: pADH100Cau
FEATURES             Location/Qualifiers
     source          1..5047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(127..984)
                     /codon_start=1
                     /label=AmpR
                     /note="color: #ff6600"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1181..1208
                     /label=AHO1096
                     /note="color: black; sequence: 
                     GACGGCACGGCCACGCGTTTAAACCGCC; added: 2024-12-10"
     misc_feature    1209..1930
                     /label=NAT 2of2
                     /note="color: #ff6600"
     primer_bind     1911..1949
                     /label=Fragment 1.FOR
                     /note="color: black; sequence: 
                     gtcaatcgtatgtgaatgctACAGACTCAATCAACGAAG"
     misc_feature    1931..2492
                     /label=CauSNR52p
                     /note="color: #ff00ff"
     primer_bind     1931..1949
                     /label=Primer 1
                     /note="color: black; sequence: 
                     ACGAGGCAAGCTTGATGTGCGGCCGCACAGACTCAATCAACGAAG"
     primer_bind     1931..1949
                     /label=SNR52p_FWD
                     /note="color: black; sequence: 
                     ACGAGGCAAGCTTGATGTGCGGCCGCACAGACTCAATCAACGAAG"
     primer_bind     1931..1949
                     /label=Fragment 1.FOR
                     /note="color: black; sequence: 
                     ACGAGGCAAGCTTGATGTGCGGCCGCACAGACTCAATCAACGAAG"
     primer_bind     2084..2103
                     /label=SNR52_F
                     /note="color: black; sequence: AAGCTTCCCTCAGATCAGGC; added:
                     2024-12-10"
     primer_bind     complement(2468..2492)
                     /label=pADH100_Cau_fgmA_R
                     /note="color: black; sequence: CTGTTTTCTGCTGAGGGAGTCGAAC; 
                     added: 2024-12-10"
     primer_bind     2473..2529
                     /label=Primer 2
                     /note="color: black; sequence: 
                     ACTCCCTCAGCAGAAAACAGGATTGAGCACGTTGACGTGGGTTTTAGAGCTAGAAATAG
                     "
     primer_bind     2473..2529
                     /label=gRNA_ade2F
                     /note="color: black; sequence: 
                     ACTCCCTCAGCAGAAAACAGGATTGAGCACGTTGACGTGGGTTTTAGAGCTAGAAATAG
                     ; added: 2024-12-10"
     primer_bind     complement(2474..2511)
                     /label=SNR52p_REV
                     /note="color: black; sequence: 
                     CGAGACGGAATTCCGTCTCCTGTTTTCTGCTGAGGGAG"
     primer_bind     complement(2474..2511)
                     /label=Fragment 1.REV
                     /note="color: black; sequence: 
                     CGAGACGGAATTCCGTCTCCTGTTTTCTGCTGAGGGAG"
     misc_RNA        2511..2599
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
                     /note="color: #00ccff; direction: RIGHT"
     primer_bind     complement(2656..2676)
                     /label=sgRNA_polyT_FRT_REV
                     /note="color: black; sequence: ATAGGAACTTCACGCGGTGGC"
     primer_bind     complement(2656..2676)
                     /label=Fragment 2.REV
                     /note="color: black; sequence: ATAGGAACTTCACGCGGTGGC"
     misc_feature    2666..2699
                     /label=FRT
                     /note="color: #000000; direction: RIGHT"
     primer_bind     complement(2685..2713)
                     /label=Fragment 1.REV
                     /note="color: black; sequence: 
                     attttgatcgGCGGGAAGTTCCTATTCTC"
     primer_bind     2694..2727
                     /label=Fragment 2.FOR
                     /note="color: black; sequence: 
                     aacttcccgcCGATCAAAATGTCTAGAGAAGATC"
     misc_feature    2703..4238
                     /label=CauHIS1_DS
                     /note="color: #ff99cc; direction: RIGHT"
     primer_bind     complement(4219..4236)
                     /label=pADH100Cau_fgmC_R
                     /note="color: black; sequence: AGCAAACGTAACGACGCG; added: 
                     2024-12-10"
     primer_bind     complement(4221..4258)
                     /label=Fragment 2.REV
                     /note="color: black; sequence: 
                     agcgagtcagtgagcgaggaCCAGCAAACGTAACGACG"
     primer_bind     complement(4234..4260)
                     /label=pADH100Cau_fgmB_R
                     /note="color: black; sequence: GCAGCGAGTCAGTGAGCGAGGACCAGC;
                     added: 2024-12-10"
     rep_origin      complement(4412..5000)
                     /direction=LEFT
                     /label=ori
                     /note="color: #ffff00"
ORIGIN
        1 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
       61 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
      121 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      181 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
      241 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
      301 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
      361 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
      421 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
      481 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
      541 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
      601 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
      661 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
      721 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
      781 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
      841 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      901 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      961 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1021 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1081 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     1141 gacattaacc tataaaaata ggcgtatcac gaggcccttt gacggcacgg ccacgcgttt
     1201 aaaccgccag gtgatgctga agctattgaa gctttggatg gttctttcac taccgatact
     1261 gtttttagag ttactgctac tggtgatggt ttcactttga gagaagttcc agttgatcca
     1321 ccattgacta aggttttccc agatgatgaa tccgatgatg aatccgatgc tggtgaagat
     1381 ggtgatccag attctagaac tttcgttgct tatggtgatg atggtgattt ggctggtttc
     1441 gttgttgttt cttattctgg ttggaacaga agattgactg ttgaagatat tgaagttgct
     1501 ccagaacata gaggtcatgg tgttggtaga gctttgatgg gtttggctac tgaattcgcc
     1561 agagaaagag gtgctggtca tttgtggttg gaagttacca atgttaatgc tccagctatt
     1621 catgcttata gaagaatggg tttcactttg tgtggtttgg atactgcttt atacgatggt
     1681 actgcttccg atggtgaaca agctttgtat atgtctatgc catgtccata ataactcgag
     1741 tcagtactga caataaaaag attcttgttt tcaagaactt gtcatttgta tagttttttt
     1801 atattgtagt tgttctattt taatcaaatg ttagcgtgat ttatattttt tttcgcctcg
     1861 acatcatctg cccagatgcg aagttaagtg cgcagaaagt aatatcatgc gtcaatcgta
     1921 tgtgaatgct acagactcaa tcaacgaagt tttttttttt tttttttttt tttttttctt
     1981 cttcatagac aataacctta tcaaagttta taaaactata tcgcaataga aaacaaaccc
     2041 ccaatcagct taacttttag agtcacttac cctacaaagg tgtaagcttc cctcagatca
     2101 ggcggcacgc atctgatatc taacatccat tgtaaaatta gtgaagatta acactcagag
     2161 ccagttattt tgaacacaga tggcttttgg ctgctcttat cgtctacgtt tctttttttt
     2221 tttttttttt tttttttttt ttttttccta attttttacg agttcactaa ttggtttcgg
     2281 gggctcaatt ttcgaatgcg acattcataa actttgacct tttatgtgtt gatgcggata
     2341 attttgacct caaaactcgg aaaaattgat gtatagcgta ttcgacatgg ctttaaaagt
     2401 cgcagagttc tacgattttc atttgcactg gcccatccgg taagcctcgg ggctccggca
     2461 tgtgagggtt cgactccctc agcagaaaac aggagacgga attccgtctc gttttagagc
     2521 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     2581 cggtgctttt tttctcgagt ttttttatcg agtgtttaag gataatgata actgaagaga
     2641 agaattagtt ttgccgccac cgcgtgaagt tcctatactt tctagagaat aggaacttcc
     2701 cgccgatcaa aatgtctaga gaagatccta ttaaggctga aaaagatttt tctgaaactc
     2761 ttgaagagca gatccctctc attgagaaga cttctgatta caaacaagct atcgagaagt
     2821 acttggtgtt ggaaaagcaa atccgtcaat cttcagattt ggctctgagt aagaagatct
     2881 taagcaaaat tgttgatacg ctagtccata atgatgattg ggactacttg gatgaattaa
     2941 tcacgccttt gtcaaagaaa catggccaac tcaagagctc cattcaactg atgattcaag
     3001 agatcatcaa gagcctcgaa acgttagata acgacaagcc tgtgcaatta gagaagaaga
     3061 tgaagattat tgaaactatt cgtacggtga ctgataagaa gatctttgtg gaggttgaga
     3121 gggcggtcgt ttccaagatg ttggctgaaa tatacttgga gaagaagaat gacttggata
     3181 aggccgcaga tatcttatgt gacttgcagg tcgagacata ctcgttgatg ccttttgaag
     3241 agaaaattga gtacattctc gagcaaatta ggttgacatt gcaaaaaaaa gactttgcac
     3301 aagcaaagat tttgagcaga aagattctct taaaaacgtt gaaaaatttt gaaagggccg
     3361 acatatttaa ggcaacgtac ttgaagtact tactagagat caacaaacat gagaatgact
     3421 acatcatggt tgtcaacaac tcacttctct tgatagagat cccactagtc aaggaatctt
     3481 cagaatacaa ggacttgctt gtgtccatca tctactacat cattctatcg ccattcgata
     3541 accttcaatc cgatttgatt cataaaatca agctcaacac tgctttttct aagaatgtgg
     3601 attctaaggt gttcaagctt ttggaaattt tcacaacaaa tgagctcatt cactctgaaa
     3661 atatcgaaat ggaatatcgt gaaaactatc ttaataagtc acctgttttt gctgataacg
     3721 agacgaacaa cacaaactta cagaacagaa ttattgagca caatttgcgt gtaatcaaca
     3781 tgtactatct gtttatcaag ttagatcggt tggctcattt gttacaagtc agttcacaag
     3841 atgcggaggc acatgtcagt ggtatggtaa acaaaggtat gatcaccgcg aaaatcaacc
     3901 gtccccaggg tattatcaag ttcgacaact tgaaacatgg tactagtgcc ggcgctgctc
     3961 agattaccag taatagtgag aacatcaatg atttgttgaa caactggata tctgacgttg
     4021 acaagctatt ggaagaggtc gacagcattg gtcacttgat taacaaggag gaaatgatgt
     4081 atggtattaa gcaaaacctg tgatatacat aatgaatagc tcaaatctta acagaatttc
     4141 ttgacccttt acttacacaa taatagcaaa aatgaaagca cccaaagagg caaatcctga
     4201 aagtgtgctc atctgattcg cgtcgttacg tttgctggtc ctcgctcact gactcgctgc
     4261 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4321 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4381 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4441 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4501 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4561 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4621 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4681 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4741 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4801 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     4861 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4921 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4981 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5041 ggaacga
//
