LOCUS       Exported                2623 bp ds-DNA     circular SYN 08-FEB-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2623)
  AUTHORS   Letizia
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Feb 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2623
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     1..24
                     /label=bbKan_Fw
     primer_bind     complement(69..90)
                     /label=Seq_pGABlue_Rv
     primer_bind     complement(120..145)
                     /label=pGA_blue_seq_2_rv
     CDS             150..965
                     /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
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(584..601)
                     /label=bbKan_Rv_interno
     primer_bind     594..610
                     /label=bbKan_Fw_interno
     rep_origin      1050..1638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1197
                     /label=Base pair substitution CG to TA
                     /note="Base pair substitution from CG to AT, confirmed by 2
                     sequencings (on the same strand, different primers). It 
                     does not seem to affect replication or plasmid yield in 
                     DH5-alpha. It can be considered silent.P.S. After a BLAST 
                     search, both versions of ori can be found: it may be an 
                     error of the original map."
     primer_bind     complement(1212..1233)
                     /label=ADAP_FL_GG_Rv
     primer_bind     1746..1765
                     /label=pGA_blue_seq_2_fw
     primer_bind     1810..1829
                     /label=Seq_pGABlue_Fw
     primer_bind     complement(1849..1868)
                     /label=bbKan_Rv
     primer_bind     1865..1892
                     /label=GFP+linker_HI_Fw
     primer_bind     1866..1928
                     /label=GFPlinker_CD_Rv
     primer_bind     1866..1927
                     /label=GFPlinker_HI_Fw
     CDS             1873..1902
                     /codon_start=1
                     /label=Linker
                     /translation="TGGSGGSSGS"
     misc_feature    1873..1876
                     /label=H
     CDS             1903..2616
                     /codon_start=1
                     /product="Aequoria victoria green fluorescent protein"
                     /label=GFP_BsaFree
                     /note="color: #05fd14"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(join(2593..2623,1..4))
                     /label=GFP_HI_GG_Rv
     primer_bind     complement(join(2595..2623,1..4))
                     /label=GFP_HI_Rv
     misc_feature    2617..2620
                     /label=I
ORIGIN
        1 gaccctgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
       61 atttaaaagg atctaggtga agatcctttt tgataatctc atgaacaata aaactgtctg
      121 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct
      181 ctaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg
      241 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag
      301 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca
      361 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc
      421 ctgatgatgc atggttactc accactgcga tccccgggaa aacagcattc caggtattag
      481 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt
      541 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgactcgctc
      601 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta
      661 atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca ttctcaccgg
      721 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat
      781 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca
      841 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat
      901 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt
      961 tctaagaatt aattcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     1021 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     1081 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     1141 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     1201 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     1261 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     1321 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     1381 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     1441 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     1501 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     1561 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     1621 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     1681 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     1741 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     1801 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     1861 attctggtct ccaccggtgg ttctggtggc tcgagtggtt caagtaaagg agaagaactt
     1921 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
     1981 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
     2041 tgcactactg gaaaactacc tgttccatgg ccaacacttg tcactacttt cacttatggt
     2101 gttcaatgct tttcaagata cccagatcat atgaaacggc atgacttttt caagagtgcc
     2161 atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg gaactacaag
     2221 acacgtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt
     2281 attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca
     2341 cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttaa cttcaaaatt
     2401 agacacaaca ttgaagatgg aagcgttcaa ctagcagacc attatcaaca aaatactcca
     2461 attggcgatg gccctgtcct tttaccagac aaccattacc tgtccacaca atctgccctt
     2521 tcgaaagatc ccaacgaaaa gagggaccac atggtccttc ttgagtttgt aacagctgct
     2581 gggattacac atggcatgga tgaactatac aaataaatag aga
//
