LOCUS       Exported                8988 bp DNA     circular SYN 27-SEP-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8988)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 27, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8988
                     /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"
     misc_feature    3604..3609
                     /label=SpeI
     misc_feature    3616..3738
                     /label=CreN59
     misc_feature    3739..3744
                     /label=KpnI
     CDS             3745..4194
                     /codon_start=1
                     /label=nMag
                     /translation="HTLYAPGGYDIMGYLDQIGNRPNPQVELGPVDTSCALILCDLKQK
                     DTPIVYASEAFLYMTGYSNAEVLGRNCRFLQSPDGMVKPKSTRKYVDSNTINTMRKAID
                     RNAEVQVEVVNFKKNGQRFVNFLTMIPVRDETGEYRYSMGFQCETE"
     CDS             4213..4233
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4249..4305
                     /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"
     misc_feature    complement(4306..4311)
                     /label=MluI
     CDS             4312..5019
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     misc_feature    complement(5023..5028)
                     /label=EcoRI
     misc_feature    5039..5044
                     /label=XhoI
     misc_feature    complement(5072..5077)
                     /label=BamHI
     misc_feature    5104..5677
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             5710..6309
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     misc_feature    complement(6323..6328)
                     /label=XhoI
     misc_feature    6441..6446
                     /label=KpnI
     LTR             6512..6745
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    complement(6877..6882)
                     /label=XbaI
     polyA_signal    6902..7126
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    complement(7163..7168)
                     /label=XhoI
     promoter        7222..7326
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7327..8187
                     /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      8358..8946
                     /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 tggccacctc tgatgaagtc aggaagaacc tgatggacat gttcagggac
     3661 aggcaggcct tctctgaaca cacctggaag atgctcctgt ctgtgtgcag atcctgggct
     3721 gcctggtgca agctgaacgg tacccatact ctttatgccc ccggtggata tgacattatg
     3781 ggatatctgg accagatcgg caaccggcca aacccgcagg tggaactggg ccccgtggat
     3841 acatcctgcg ccttgattct ttgtgacctg aaacagaaag acaccccgat agtttacgcg
     3901 agtgaagcct tcctctacat gacaggttac agcaacgcag aggtgctggg ccggaattgc
     3961 cggtttctgc aaagccctga cggcatggtg aagcccaaga gcacccggaa gtacgtggat
     4021 agtaacacaa tcaatactat gcgcaaggca atcgacagga atgccgaggt gcaggttgaa
     4081 gtagtcaatt ttaaaaagaa tggacagcga tttgttaatt tcctgactat gatacctgtt
     4141 agggacgaaa caggcgagta tcgatactct atgggattcc agtgcgaaac agaaggcgga
     4201 agcggtggcg tgcccaagaa gaagaggaaa gtcggatccg gcagcggcgc caccaacttc
     4261 agcctgctga agcaggccgg cgacgtggag gagaaccccg gccccacgcg tgtgagcaag
     4321 ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     4381 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     4441 ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     4501 atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc
     4561 cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc
     4621 gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc
     4681 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     4741 accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc
     4801 gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc
     4861 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg
     4921 gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     4981 cgccactcca ccggcggcat ggacgagctg tacaagtaat aggaattcga agcttgagct
     5041 cgagatctga gtccggtagc gctagttcga aggatccgca tgcatctagg gcggccaatt
     5101 ccgcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     5161 gtgtgcgttt gtctatatgt gattttccac catattgccg tcttttggca atgtgagggc
     5221 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     5281 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag
     5341 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     5401 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     5461 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     5521 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     5581 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     5641 cggggacgtg gttttccttt gaaaaacacg atgataagct tgccacaacc cacaaggaga
     5701 cgaccttcca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     5761 cgggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     5821 gacccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     5881 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     5941 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     6001 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     6061 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     6121 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     6181 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     6241 gtcaccgccg acgtcgagtg cccgaaggac cgcgcgacct ggtgcatgac ccgcaagccc
     6301 ggtgcctgat aataggcggc cgctcgagac ctagaaaaac atggagcaat cacaagtagc
     6361 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     6421 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     6481 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     6541 caagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     6601 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     6661 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     6721 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     6781 taacttgcaa agaaatgaat atcagagagt gagaggcctt gacattataa tagatttagc
     6841 aggaattgaa ctaggagtgg agcacacagg caaagttcta gagctcgctg atcagcctcg
     6901 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     6961 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     7021 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     7081 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     7141 agaaccagct gggggcgcgc ccctcgaggc cgccatggtc atagctgttt gacgtcaggt
     7201 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7261 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7321 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7381 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7441 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7501 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7561 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7621 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7681 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7741 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7801 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7861 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7921 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7981 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     8041 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     8101 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8161 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8221 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8281 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8341 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8401 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8461 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     8521 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8581 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8641 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8701 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc
     8761 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8821 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8881 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8941 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgc
//
