LOCUS       pF_AA414        3607 bp DNA     circular SYN 14-MAR-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3607)
  AUTHORS   GenScript
  TITLE     Direct Submission
  JOURNAL   Exported Dec 21, 2023 from SnapGene 7.1.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    432..443
                     /label=GG-2F
     CDS             457..459
                     /label=Start
     CDS             460..501
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5_v1.1
                     /translation="GKPIPNPLLGLDST"
     CDS             502..1389
                     /codon_start=1
                     /product="HaloTag(R), a modified bacterial dehalogenase 
                     that forms covalent bonds with chloroalkane derivatives
                     
                     Addgene 185762"
                     /label=HaloTag_v1.1
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     misc_difference 1327..1329
                     /label=GAC->GAT
     misc_difference 1390..1395
                     /label=MluI site
                     /note="From most maps of degron panel described by Bondeson
                     et al, Nat Commun. 2022. PMID: 36127368
                     Originally included by Bondeson et al as a MluI site for 
                     digestion verification purposes; preserved to maintain 
                     consistency, as it is in a coding region.
                     
                     "
     CDS             1396..1446
                     /product="Rigid linker used by Bondeson et al, Nat Commun. 
                     2022. PMID: 36127368
                     Original linker sequence (19 residues, AEAAAKEAAAKEAAAKAGS)
                     includes GS at 3' end; to avoid redundancy, removed GS 
                     because GPP's Module 3 Golden Gate overhangs already code 
                     for GS.
                     
                     "
                     /label=Degron-linker_v1.1
     misc_feature    1447..1459
                     /label=GG-3R
     CDS             1447..1452
                     /codon_start=1
                     /label=GS
                     /translation="GS"
     rep_origin      complement(1919..2507)
                     /direction=LEFT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2685..3494)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=aph(3')-Ia
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagagaatt cgagctcggt acctcgcgaa
      421 tacatctaga tgaagacgta ggagagagcc gccaccatgg gtaagcctat ccctaaccct
      481 ctcctcggtc tcgattctac ggcagaaatc ggtactggct ttccattcga cccccattat
      541 gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga tggcacccct
      601 gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat catcccgcat
      661 gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa atccgacaaa
      721 ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt catcgaagcc
      781 ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct gggtttccac
      841 tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt catccgccct
      901 atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc cttccgcacc
      961 accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg tacgctgccg
     1021 atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga gccgttcctg
     1081 aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat cgccggtgag
     1141 ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca gtcccctgtc
     1201 ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga agccgctcgc
     1261 ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct gaatctgctg
     1321 caagaagata acccggacct gatcggcagc gagatcgcgc gctggctgtc gacgctcgag
     1381 atttccggca cgcgtgctga agctgctgca aaggaagctg cagctaagga ggctgcagct
     1441 aaggctggca gcagtcttca tcggatcccg ggcccgtcga ctgcagaggc ctgcatgcaa
     1501 gcttggtgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     1561 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     1621 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     1681 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1741 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1801 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1861 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2521 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2581 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2641 caagcccaat ctgaataatg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg
     2701 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     2761 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     2821 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     2881 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     2941 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     3001 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     3061 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     3121 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     3181 aatgctgttt ttccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     3241 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     3301 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     3361 ggcttcccat acaagcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     3421 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga cgtttcccgt
     3481 tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt
     3541 catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga cacgggccag
     3601 agctgca
//
