LOCUS       sequence-13844-e        2882 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 2882)
  TITLE     2-series polycistronic destination vectors
REFERENCE   2  (bases 1 to 2882)
  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: pET empty polycistronic destination vector (2E)
FEATURES             Location/Qualifiers
     source          1..2882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     102..119
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     103..119
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(333..351)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      347..394
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(529..1344)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSGRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1252..1271
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1336..1355
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      1466..2054
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1955..1974
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2208..2225
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2240..2379
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2305..2324)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2465..2487
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2481..2672)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggcaccatgc atcgatcccg cgaaattaat acgactcact atagggagac cacaacggtt
       61 tccctctaga aggcctcgga tccgggccct gtacaagatc ctgtaaaacg acggccagtg
      121 aattcactcg agtcccgggg cgatcgcagc ggataacaat ttcacatcct gcagggatat
      181 cggcgcgccc gccagggttt tcccagtcgc ggccgctcgc gcacctggtg ccatcgccct
      241 gatagacgtt aattaaagta ctggccggcc acgcgtctcg agagatccgg ctgctaacaa
      301 agcccgaaag gaagctgagt tggctgctgc caccgctgag caataactag cataacccct
      361 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccgacta
      421 gtaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
      481 ttatttttct aaatacattc aaatatgtat ccgctcatga attaattctt agaaaaactc
      541 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
      601 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
      661 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
      721 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
      781 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
      841 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
      901 acgaaatacg cgaccgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
      961 caggaacact gccagcgcat caacaatgtt ttcacctgaa tcaggatatt cttctaatac
     1021 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     1081 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     1141 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     1201 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     1261 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tagagcaaga
     1321 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     1381 ttttattgtt catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     1441 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     1501 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     1561 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     1621 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     1681 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     1741 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     1801 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     1861 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     1921 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     1981 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     2041 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     2101 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     2161 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     2221 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     2281 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     2341 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     2401 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     2461 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     2521 ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc
     2581 gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc
     2641 atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg gggatttctg
     2701 ttcatggggt aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg
     2761 atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc
     2821 gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg
     2881 tt
//
