LOCUS       sequence-9227-ex        3174 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3174)
  TITLE     Forth generation Seydoux lab vectors
REFERENCE   2  (bases 1 to 3174)
  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"
COMMENT     Sequence Label: pCM1.59
FEATURES             Location/Qualifiers
     source          1..3174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1219..1343
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1415..1434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1416..1434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1439..1455)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1439..1455)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1557..1576)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1568..2377
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1635..1654)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2524..3112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3013..3032
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttgat tgagatacac aagtgttttt tatttatttt
      721 cctgtttaaa taaattcgat tcattttata aaactctgaa ttgaaagtat tcaattaaat
      781 tctgaaaagc ctcatgcaaa tcagcgcacg gcttaatagg cgtggccaat tcctgccgtt
      841 tcgccacccg tgcgcgcatt tctggaactt aaaggaacat gttgcaccgc gacgtatggg
      901 tctcgcagcg atcgcaaaca ggtatacggt aaaagtttcg aaatttagga cttcacctca
      961 aaacttttta gagttttatt ttttcgatgc tttttggttg ttttatattt cgtttcatct
     1021 aaaaatttta gaaataatta tttctatttg aatatgacca ctttcacatt ttcctcgaat
     1081 gtttcattcc gaaattctag tgaatattcg ttagataagc tagatgagcg aatcgccgca
     1141 gagtttaagc ttatgtcgtt acgttattga aaaatatatt attactaagt tttcttacag
     1201 gttaaccatc gaagaatgac aagtttgtac aaaaaagttg aacgagaaac gtaaaatgat
     1261 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     1321 acacaacata tgcagtcact atgaatcaac tacttagatg gtattagtga cctgtagaat
     1381 tcgagctcta gagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt
     1441 catagctgtt tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca
     1501 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag
     1561 gggtgttatg agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga
     1621 tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat
     1681 ctatcgcttg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag
     1741 cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc
     1801 tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc
     1861 gatccccgga aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat
     1921 tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc
     1981 ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt
     2041 ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa
     2101 agaaatgcat aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc
     2161 acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt
     2221 cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc
     2281 tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa
     2341 attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta
     2401 acactggcag agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct
     2461 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2521 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2581 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2641 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2701 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2761 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2821 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2881 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     2941 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3001 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3061 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3121 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
