LOCUS       5'_entry_clone_4        2743 bp DNA     circular SYN 22-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2743)
  AUTHORS   Marleen Aarts
  TITLE     Direct Submission
  JOURNAL   Exported Jan 22, 2024 from SnapGene 6.2.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      137..164
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      256..342
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     391..413
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     405..422
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     406..422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    438..537
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    576..589
                     /label=PRE
     misc_feature    603..616
                     /label=PRE
     misc_feature    630..643
                     /label=PRE
     misc_feature    657..670
                     /label=PRE
     misc_feature    677..707
                     /label=2YBTATA
     protein_bind    complement(727..822)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(853..872)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(854..872)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(877..893)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(877..893)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(995..1014)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1006..1815
                     /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(1073..1092)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1962..2550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2451..2470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2704..2721
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
      121 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
      181 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
      241 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
      301 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      361 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      421 gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
      481 aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctgga
      541 gacccaagct tggtaccgag ctcggatctg tagctagaac atcctgtaca gtatccgtag
      601 ctagaacatc ctgtacagga tctgtagcta gaacatcctg tacagtatcc gtagctagaa
      661 catcctgtac aggatcctct agagggtata taatgggggc cactcgagca tgcatctaga
      721 ggcacccaac ttttctatac aaagttggca ttataagaaa gcattgctta tcaatttgtt
      781 gcaacgaaca ggtcactatc agtcaaaata aaatcattat ttgccatcca gctgcagggc
      841 ggccgcgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
      901 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
      961 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1021 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1081 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1141 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1201 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1261 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     1321 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     1381 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     1441 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     1501 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     1561 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     1621 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     1681 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     1741 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     1801 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     1861 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     1921 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1981 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2041 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2101 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2161 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2221 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2281 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2341 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2401 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2461 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2521 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2581 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2641 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2701 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tac
//
