LOCUS       Exported                7270 bp ds-DNA     circular SYN 12-MAR-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pAK026_pHAGE-EFS-MCP-YPet
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7270)
  AUTHORS   Jeltsch Group
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jan 29, 2024 from SnapGene 3.3.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7270
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2058..2175
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2265..2476
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      2522..2531
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2528..2878
                     /codon_start=1
                     /label=MCP
                     /translation="MASNFTQFVLVDNGGTGDVTVAPSNFANGIAEWISSNSRSQAYKV
                     TCSVRQSSAQNRKYTIKVEVPKGAWRSYLNMELTIPIFATNSDCELIVKAMQGLLKDGN
                     PIPSAIAANSGIY"
     CDS             2906..3622
                     /codon_start=1
                     /product="yellow fluorescent protein optimized for FRET 
                     with CyPet (Nguyen and Daugherty, 2005)"
                     /label=YPet
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLLCTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDSNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL
                     EFLTAAGITEGMNELYK"
     misc_feature    3308
                     /label=G to A
     misc_feature    3499
                     /label=C to T
     misc_feature    3598
                     /label=C to A
     CDS             3629..3649
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     misc_feature    3660..4248
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(4131..4142)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4323..4556
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     promoter        4761..4865
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4866..5726
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5897..6485
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6896..6917
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6932..6962
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6970..6986
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     6994..7010
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg
     2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa
     2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca
     2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat
     2221 gctagctgat gcgggccctt cgagtggctc cggtgcccgt cagtgggcag agcgcacatc
     2281 gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag
     2341 gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg
     2401 tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt
     2461 tgccgccaga acacaggtgt cgtgacgcgg ttccggttaa ggtgaacgcg tgcaggctgg
     2521 cgccaccatg gcttctaact ttactcagtt cgttctcgtc gacaatggcg gaactggcga
     2581 cgtgactgtc gccccaagca acttcgctaa cgggatcgct gaatggatca gctctaactc
     2641 gcgttcacag gcttacaaag taacctgtag cgttcgtcag agctctgcgc agaatcgcaa
     2701 atacaccatc aaagtcgagg tgcctaaagg cgcctggcgt tcgtacttaa atatggaact
     2761 aaccattcca attttcgcca cgaattccga ctgcgagctt attgttaagg caatgcaagg
     2821 tctcctaaaa gatggaaacc cgattccctc agcaatcgca gcaaactccg gcatctacgg
     2881 gggatctggt agcggcgctg gatccatggt gagcaaaggc gaagagctgt tcaccggcgt
     2941 ggtgcccatc ctggtggagc tggacggcga cgtgaacggc cacaagttca gcgtgagcgg
     3001 cgagggcgag ggcgacgcca cctacggcaa gctgaccctg aagctgctgt gcaccaccgg
     3061 caagctgccc gtgccctggc ccaccctggt gaccaccctg ggctacggcg tgcagtgctt
     3121 cgcccggtac cccgaccaca tgaagcagca cgacttcttc aagagcgcca tgcccgaggg
     3181 ctacgtgcag gagcggacca tcttcttcaa ggacgacggc aactacaaga cccgggccga
     3241 ggtgaagttc gagggcgaca ccctggtgaa ccggatcgag ctgaagggca tcgacttcaa
     3301 ggaggacagc aacatcctgg gccacaagct ggagtacaac tacaacagcc acaacgtgta
     3361 catcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc ggcacaacat
     3421 cgaggacggc ggcgtgcagc tggccgacca ctaccagcag aacaccccca tcggcgacgg
     3481 ccccgtgctg ctgcccgata accactacct gagctaccag agcgccctgt tcaaggaccc
     3541 caacgagaag cgggaccaca tggtgctgct ggagttcctg accgccgccg gcatcacaga
     3601 gggcatgaac gagctgtata agctcgagcc aaagaaaaag cggaaagtgt aatctagata
     3661 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3721 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3781 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3841 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3901 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3961 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     4021 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     4081 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     4141 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     4201 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct gagatccttt
     4261 aagaccaatg acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg
     4321 actggaaggg ctaattcact cccaacgaag acaagatctg ctttttgctt gtactgggtc
     4381 tctctggtta gaccagatct gagcctggga gctctctggc taactaggga acccactgct
     4441 taagcctcaa taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga
     4501 ctctggtaac tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag
     4561 tagttcatgt catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag
     4621 tgagaggccc gggttaatta aggaaagggc tagatcattc ttgaagacga aagggcctcg
     4681 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4741 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4801 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4861 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4921 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4981 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5041 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5101 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5161 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5221 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5281 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5341 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5401 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5461 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5521 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5581 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5641 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5701 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5761 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5821 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5881 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5941 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6001 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6061 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6121 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6181 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6241 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6301 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6361 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6421 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6481 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6541 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6601 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6661 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6721 gcaagctcat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     6781 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     6841 ccccgtggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     6901 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     6961 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac atgattacga
     7021 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     7081 atgtatctta tcatgtctgg atcaactgga taactcaagc taaccaaaat catcccaaac
     7141 ttcccacccc ataccctatt accactgcca attacctgtg gtttcattta ctctaaacct
     7201 gtgattcctc tgaattattt tcattttaaa gaaattgtat ttgttaaata tgtactacaa
     7261 acttagtagt
//
