LOCUS       plenti-FLAG-CAMK        8536 bp DNA     circular SYN 22-MAY-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8536)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 22, 2024 from SnapGene 7.2.0
            https://www.snapgene.com
COMMENT     Sequence Label: plenti-FLAG-CAMK2D
FEATURES             Location/Qualifiers
     source          1..8536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..816
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1683..1916
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2101..2145
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2443..2560
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        2643..3022
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3023..3226
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        3252..3270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4828..4857
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             4876..4899
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4918..4974
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             4975..5574
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     protein_bind    5631..5664
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    5720..6308
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(6191..6202)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6311..6327)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6837..7017
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7079..7667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7744..8403)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(8404..8508)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gctctagaca tgtccaatat gaccgccatg
     2641 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     2701 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     2761 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     2821 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     2881 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     2941 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     3001 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     3061 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     3121 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     3181 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     3241 tcagaatttt gtaatacgac tcactatagg gcggccggga attcgtcgac tggatccggt
     3301 accgaggaga tctgccgccg cgatcgcatg gcttcgacca caacctgcac caggttcacg
     3361 gacgagtatc agcttttcga ggagcttgga aagggggcat tctcagtggt gagaagatgt
     3421 atgaaaattc ctactggaca agaatatgct gccaaaatta tcaacaccaa aaagctttct
     3481 gctagggatc atcagaaact agaaagagaa gctagaatct gccgtctttt gaagcaccct
     3541 aatattgtgc gacttcatga tagcatatca gaagagggct ttcactactt ggtgtttgat
     3601 ttagttactg gaggtgaact gtttgaagac atagtggcaa gagaatacta cagtgaagct
     3661 gatgccagtc attgtataca gcagattcta gaaagtgtta atcattgtca cctaaatggc
     3721 atagttcaca gggacctgaa gcctgagaat ttgcttttag ctagcaaatc caagggagca
     3781 gctgtgaaat tggcagactt tggcttagcc atagaagttc aaggggacca gcaggcgtgg
     3841 tttggttttg ctggcacacc tggatatctt tctccagaag ttttacgtaa agatccttat
     3901 ggaaagccag tggatatgtg ggcatgtggt gtcattctct atattctact tgtggggtat
     3961 ccacccttct gggatgaaga ccaacacaga ctctatcagc agatcaaggc tggagcttat
     4021 gattttccat caccagaatg ggacacggtg actcctgaag ccaaagacct catcaataaa
     4081 atgcttacta tcaaccctgc caaacgcatc acagcctcag aggcactgaa gcacccatgg
     4141 atctgtcaac gttctactgt tgcttccatg atgcacagac aggagactgt agactgcttg
     4201 aagaaattta atgctagaag aaaactaaag ggtgccatct tgacaactat gctggctaca
     4261 aggaatttct cagcagccaa gagtttgttg aagaaaccag atggagtaaa ggagtcaact
     4321 gagagttcaa atacaacaat tgaggatgaa gatgtgaaag cacgaaagca agagattatc
     4381 aaagtcactg aacaactgat cgaagctatc aacaatgggg actttgaagc ctacacaaaa
     4441 atctgtgacc caggccttac tgcttttgaa cctgaagctt tgggtaattt agtggaaggg
     4501 atggattttc accgattcta ctttgaaaat gctttgtcca aaagcaataa accaatccac
     4561 actattattc taaaccctca tgtacatctg gtaggggatg atgccgcctg catagcatat
     4621 attaggctca cacagtacat ggatggcagt ggaatgccaa agacaatgca gtcagaagag
     4681 actcgtgtgt ggcaccgccg ggatggaaag tggcagaatg ttcattttca tcgctcgggg
     4741 tcaccaacag tacccatcaa gccaccctgt attccaaatg ggaaagaaaa cttctcagga
     4801 ggcacctctt tgtggcaaaa catcctcgag cagaaactca tctcagaaga ggatctggca
     4861 gcaaatgata tcctggatta caaggatgac gacgataagg tttgggtagg aagcggagct
     4921 actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctatgacc
     4981 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     5041 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggatcgccac
     5101 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     5161 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     5221 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     5281 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     5341 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     5401 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     5461 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     5521 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatgtaca
     5581 cccaaacggc cggccgcggt ctgtacaagt aggattcgtc gagggaccta ataacttcgt
     5641 atagcataca ttatacgaag ttatacatgt ttaagggttc cggttccact aggtacaatt
     5701 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     5761 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     5821 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     5881 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     5941 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     6001 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     6061 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     6121 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     6181 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     6241 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     6301 ctccccgcat cgataccgtc gacctcgatc gagacctaga aaaacatgga gcaatcacaa
     6361 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg
     6421 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg
     6481 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
     6541 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattggc
     6601 agaactacac accagggcca gggatcagat atccactgac ctttggatgg tgctacaagc
     6661 tagtaccagt tgagcaagag aaggtagaag aagccaatga aggagagaac acccgcttgt
     6721 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtatta gagtggaggt
     6781 ttgacagccg cctagcattt catcacatgg cccgagagct gcatccggac tgtactgggt
     6841 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
     6901 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
     6961 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagca
     7021 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7081 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7141 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7201 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7261 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7321 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7381 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7441 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7501 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7561 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7681 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7741 tgattacgcc ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc
     7801 gacatggaag ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt
     7861 gtcgccttgc gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt
     7921 ggccacgttt aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat
     7981 attctcaata aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg
     8041 cgaatatatg tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa
     8101 cgtttcagtt tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag
     8161 ctcaccgtct ttcattgcca tacggaactc cggatgagca ttcatcaggc gggcaagaat
     8221 gtgaataaag gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt
     8281 aatatccagc tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa
     8341 atgttcttta cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc
     8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8521 aaaagtgcca cctgac
//
