LOCUS       Exported                9696 bp DNA     circular SYN 27-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pSin_EF1a_tBFP_P2A_NLS_pMag_CreC60_IRES_PuroR
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9696)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             117..750
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    797..922
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1415..1648
                     /label=RRE
     misc_feature    2164..2281
                     /label=cPPT
     promoter        2409..3587
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     CDS             3610..4308
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             4324..4380
                     /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             4390..4410
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4417..4866
                     /codon_start=1
                     /label=pMag
                     /translation="HTLYAPGGYDIMGYLRQIRNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     CDS             4873..5724
                     /codon_start=1
                     /label=CreC 60-343
                     /translation="NRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     misc_feature    5812..6385
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             6418..7017
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     LTR             7220..7453
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    7610..7834
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        7930..8034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8035..8895
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      9066..9654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cggaccgcca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
       61 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttgga
      121 agggctaatt cactcccaaa gaagacaaga tatccttgat ctgtggatct accacacaca
      181 aggctacttc cctgattagc agaactacac accagggcca ggggtcagat atccactgac
      241 ctttggatgg tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa
      301 aggagagaac accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag
      361 agaagtgtta gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct
      421 gcatccggag tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg
      481 ggactttcca gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg
      541 catataagca gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct
      601 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
      661 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
      721 ccttttagtc agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa
      781 agggaaacca gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag
      841 aggcgagggg cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga
      901 gagagatggg tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa
      961 aattcggtta aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag
     1021 cagggagcta gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag
     1081 acaaatactg ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt
     1141 atataataca gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa
     1201 ggaagcttta gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc
     1261 ggccgctgat cttcagacct ggaggaggag atatgaggga caattggaga agtgaattat
     1321 ataaatataa agtagtaaaa attgaaccat taggagtagc acccaccaag gcaaagagaa
     1381 gagtggtgca gagagaaaaa agagcagtgg gaataggagc tttgttcctt gggttcttgg
     1441 gagcagcagg aagcactatg ggcgcagcgt caatgacgct gacggtacag gccagacaat
     1501 tattgtctgg tatagtgcag cagcagaaca atttgctgag ggctattgag gcgcaacagc
     1561 atctgttgca actcacagtc tggggcatca agcagctcca ggcaagaatc ctggctgtgg
     1621 aaagatacct aaaggatcaa cagctcctgg ggatttgggg ttgctctgga aaactcattt
     1681 gcaccactgc tgtgccttgg aatgctagtt ggagtaataa atctctggaa cagatttgga
     1741 atcacacgac ctggatggag tgggacagag aaattaacaa ttacacaagc ttaatacact
     1801 ccttaattga agaatcgcaa aaccagcaag aaaagaatga acaagaatta ttggaattag
     1861 ataaatgggc aagtttgtgg aattggttta acataacaaa ttggctgtgg tatataaaat
     1921 tattcataat gatagtagga ggcttggtag gtttaagaat agtttttgct gtactttcta
     1981 tagtgaatag agttaggcag ggatattcac cattatcgtt tcagacccac ctcccaaccc
     2041 cgaggggacc cgacaggccc gaaggaatag aagaagaagg tggagagaga gacagagaca
     2101 gatccattcg attagtgaac ggatctcgac ggtatcgcca caaatggcag tattcatcca
     2161 caattttaaa agaaaggggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     2221 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     2281 tcgggtttat tacagggaca gcagagatcc actttggtcg ataagctttg caaagatgga
     2341 taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc ttgaaaggag
     2401 tgggaattgg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     2461 agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc ggggtaaact
     2521 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     2581 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg
     2641 taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc ccttgcgtgc
     2701 cttgaattac ttccacctgg ctgcagtacg tgattcttga tcccgagctt cgggttggaa
     2761 gtgggtggga gagttcgagg ccttgcgctt aaggagcccc ttcgcctcgt gcttgagttg
     2821 aggcctggcc tgggcgctgg ggccgccgcg tgcgaatctg gtggcacctt cgcgcctgtc
     2881 tcgctgcttt cgataagtct ctagccattt aaaatttttg atgacctgct gcgacgcttt
     2941 ttttctggca agatagtctt gtaaatgcgg gccaagatct gcacactggt atttcggttt
     3001 ttggggccgc gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg
     3061 gcctgcgagc gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc
     3121 tggtgcctgg cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt
     3181 cggcaccagt tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa
     3241 aatggaggac gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg
     3301 cctttccgtc ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc
     3361 acctcgatta gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt
     3421 atgcgatgga gtttccccac actgagtggg tggagactga agttaggcca gcttggcact
     3481 tgatgtaatt ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc
     3541 ctcagacagt ggttcaaagt ttttttcttc catttcaggt gtcgtgagga attgatcctt
     3601 cgaactagta tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
     3661 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     3721 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
     3781 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc
     3841 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
     3901 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     3961 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca
     4021 ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga
     4081 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca
     4141 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac
     4201 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca
     4261 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaatac gcgtggcagc
     4321 ggcgccacca acttcagcct gctgaagcag gccggcgacg tggaggagaa ccccggcccc
     4381 ctcgaggtgc ccaagaagaa gaggaaagtc ggcggacata ctctttatgc ccccggtgga
     4441 tatgacatta tgggatatct gaggcagatc aggaaccggc caaacccgca ggtggaactg
     4501 ggccccgtgg atacatcctg cgccttgatt ctttgtgacc tgaaacagaa agacaccccg
     4561 atagtttacg cgagtgaagc cttcctctac atgacaggtt acagcaacgc agaggtgctg
     4621 ggccggaatt gccggtttct gcaaagccct gacggcatgg tgaagcccaa gagcacccgg
     4681 aagtacgtgg atagtaacac aatcaatact atgcgcaagg caatcgacag gaatgccgag
     4741 gtgcaggttg aagtagtcaa ttttaaaaag aatggacagc gatttgttaa tttcctgact
     4801 atgatacctg ttagggacga aacaggcgag tatcgatact ctatgggatt ccagtgcgaa
     4861 acagaaggta ccaacaggaa atggttccct gctgaacctg aggatgtgag ggactacctc
     4921 ctgtacctgc aagccagagg cctggctgtg aagaccatcc aacagcacct gggccagctc
     4981 aacatgctgc acaggagatc tggcctgcct cgcccttctg actccaatgc tgtgtccctg
     5041 gtgatgagga gaatcagaaa ggagaatgtg gatgctgggg agagagccaa gcaggccctg
     5101 gcctttgaac gcactgactt tgaccaagtc agatccctga tggagaactc tgacagatgc
     5161 caggacatca ggaacctggc cttcctgggc attgcctaca acaccctgct gcgcattgcc
     5221 gaaattgcca gaatcagagt gaaggacatc tcccgcaccg atggtgggag aatgctgatc
     5281 cacattggca ggaccaagac cctggtgtcc acagctggtg tggagaaggc cctgtccctg
     5341 ggggttacca agctggtgga gagatggatc tctgtgtctg gtgtggctga tgaccccaac
     5401 aactacctgt tctgccgggt cagaaagaat ggtgtggctg ccccttctgc cacctcccaa
     5461 ctgtccaccc gggccctgga agggatcttt gaggccaccc accgcctgat ctatggtgcc
     5521 aaggatgact ctgggcagag atacctggcc tggtctggcc actctgccag agtgggtgct
     5581 gccagggaca tggccagggc tggtgtgtcc atccctgaaa tcatgcaggc tggtggctgg
     5641 accaatgtga acattgtgat gaactacatc agaaacctgg actctgagac tggggccatg
     5701 gtgaggctgc tcgaggatgg ggactaatag gaattcgaag cttgagctcg agatctgagt
     5761 ccggtagcgc tagttcgaag gatccgcatg catctagggc ggccaattcc gcccctctcc
     5821 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt
     5881 ctatatgtga ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg
     5941 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg
     6001 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc
     6061 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca
     6121 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag
     6181 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa
     6241 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt
     6301 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt
     6361 tttcctttga aaaacacgat gataagcttg ccacaaccca caaggagacg accttccatg
     6421 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc
     6481 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga cccggaccgc
     6541 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     6601 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     6661 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     6721 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     6781 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     6841 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     6901 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     6961 gtcgagtgcc cgaaggaccg cgcgacctgg tgcatgaccc gcaagcccgg tgcctgataa
     7021 taggcggccg ctcgagacct agaaaaacat ggagcaatca caagtagcaa tacagcagct
     7081 accaatgctg attgtgcctg gctagaagca caagaggagg aggaggtggg ttttccagtc
     7141 acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct tagccacttt
     7201 ttaaaagaaa aggggggact ggaagggcta attcactccc aacgaagaca agatctgctt
     7261 tttgcttgta ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa
     7321 ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt
     7381 gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg
     7441 aaaatctcta gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag
     7501 aaatgaatat cagagagtga gaggccttga cattataata gatttagcag gaattgaact
     7561 aggagtggag cacacaggca aagttctaga gctcgctgat cagcctcgac tgtgccttct
     7621 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     7681 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     7741 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     7801 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     7861 gggcgcgccc ctcgaggccg ccatggtcat agctgtttga cgtcaggtgg cacttttcgg
     7921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     7981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     8041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     8101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     8161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     8221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     8281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     8341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     8401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     8461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     8521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     8581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     8641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     8701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     8761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     8821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     8881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     8941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     9001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     9061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     9121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     9181 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     9241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     9301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     9361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     9421 acgacctaca ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc
     9481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     9541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     9601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     9661 agcaacgcgg cctttttacg gttcctggcc ttttgc
//
