LOCUS       Exported                8728 bp DNA     circular SYN 04-MAR-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8728)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 7, 2022 from SnapGene Viewer 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..500
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             540..1136
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     CDS             1155..1208
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1209..1910
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    2271..2495
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(2836..2854)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2875..2891)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2899..2915
                     /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)."
     promoter        complement(2923..2953)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2968..2989
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3277..3865)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4036..4896)
                     /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"
     promoter        complement(4897..5001)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5027..5482)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     5624..5640
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        5650..5668
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        5739..6042
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6043..6246
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6260..6440
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6487..6612
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    7111..7344
                     /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    7871..7988
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(8323..8341)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        complement(8393..8411)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_feature    8427..8446
                     /label=Bip recognition (reverse complement)
     promoter        complement(8456..8696)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
ORIGIN
        1 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg
       61 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc
      121 gccaaccggc tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca
      181 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
      241 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
      301 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg
      361 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct
      421 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt
      481 cctcatctcc gggcctttcg acctgcatcc atctagatct cgagcagctg aagcttacca
      541 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac
      601 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc
      661 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca
      721 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga
      781 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt
      841 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc
      901 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca
      961 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg
     1021 agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc gtcaccgccg
     1081 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgccggcg
     1141 gcgggtccgg aggagagggc agaggaagtc tcctaacatg cggtgacgtg gaggagaatc
     1201 ctggcccaat gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca
     1261 ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
     1321 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
     1381 tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg
     1441 acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgagg
     1501 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
     1561 acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac
     1621 tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg gaaggcagaa
     1681 acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc aagaccacat
     1741 atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca
     1801 gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac gaggtggcag
     1861 tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattga gcggccgcta
     1921 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
     1981 aaagggggga ctggaagggc taattcactc ccaaagaagt caagatctgc tttttgcctg
     2041 tactgggtct ctctggttag accagagtct ctctggttag accagatctg agcctgggag
     2101 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     2161 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     2221 tagtcagtgt ggaaaatctc tagcagttta aacccgctga tcagcctcga ctgtgccttc
     2281 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     2341 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     2401 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagtcaa
     2461 tagcaggcat gctggggatg cggtgggctc tatgggcggc cgttaatgat atctataaca
     2521 agaaaatata tatataataa gttatcacgt aagtagaaca tgaaataaca atataattat
     2581 cgtatgagtt aaatcttaaa agtcacgtaa aagataatca tgcgtcattt tgactcacgc
     2641 ggtcgttata gttcaaaatc agtgacactt accgcattga caagcacgcc tcacgggagc
     2701 tccaagcggc gactgagatg tcctaaatgc acagcgacgg attcgcgcta tttagaaaga
     2761 gagagcaata tttcaagaat gcatgcgtca attttacgca gactatcttt ctagggttaa
     2821 attaagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     2881 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     2941 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3001 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3061 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3121 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3181 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3241 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3301 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3361 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3421 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3481 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3541 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3601 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3661 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     3721 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3781 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3901 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     4921 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4981 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     5041 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5101 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5161 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5221 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5281 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5341 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5401 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     5461 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     5521 cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg
     5581 cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt
     5641 gagcgcgcgt aatacgactc actatagggc gaattgacta gttattaata gtaatcaatt
     5701 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     5761 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     5821 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     5881 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     5941 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     6001 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     6061 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     6121 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     6181 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     6241 agcgcgtttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc
     6301 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt
     6361 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc
     6421 agtgtggaaa atctctagca gtggcgcccg aacagggact tgaaagcgaa agggaaacca
     6481 gaggagctct ctcgacgcag gactcggctt gctgaagcgc gcacggcaag aggcgagggg
     6541 cggcgactgg tgagtacgcc aaaaattttg actagcggag gctagaagga gagagatggg
     6601 tgcgagagcg tcagtattaa gcgggggaga attagatcgc gatgggaaaa aattcggtta
     6661 aggccagggg gaaagaaaaa atataaatta aaacatatag tatgggcaag cagggagcta
     6721 gaacgattcg cagttaatcc tggcctgtta gaaacatcag aaggctgtag acaaatactg
     6781 ggacagctac aaccatccct tcagacagga tcagaagaac ttagatcatt atataataca
     6841 gtagcaaccc tctattgtgt gcatcaaagg atagagataa aagacaccaa ggaagcttta
     6901 gacaagatag aggaagagca aaacaaaagt aagaccaccg cacagcaagc ggccggccgc
     6961 gctgatcttc agacctggag gaggagatat gagggacaat tggagaagtg aattatataa
     7021 atataaagta gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt
     7081 ggtgcagaga gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc
     7141 agcaggaagc actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt
     7201 gtctggtata gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct
     7261 gttgcaactc acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag
     7321 atacctaaag gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac
     7381 cactgctgtg ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca
     7441 cacgacctgg atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt
     7501 aattgaagaa tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa
     7561 atgggcaagt ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt
     7621 cataatgata gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt
     7681 gaatagagtt aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag
     7741 gggacccgac aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc
     7801 cattcgatta gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat
     7861 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     7921 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
     7981 aaaattttcg ggtttattac agggacagca gagatccagt ttggttagta ccgggcccta
     8041 cgcgttactt aaccctagaa agataatcat attgtgacgt acgttaaaga taatcatgcg
     8101 taaaattgac gcatgtgttt tatcggtctg tatatcgagg tttatttatt aatttgaata
     8161 gatattaagt tttattatat ttacacttac atactaataa taaattcaac aaacaattta
     8221 tttatgttta tttatttatt aaaaaaaaac aaaaactcaa aatttcttct ataaagtaac
     8281 aaagcaaaaa aagcaccgac tcggtgccac ttggccctgc agacatgggt gatcctcatg
     8341 ttggccaagt tgataacgga ctagccttat tttaacttgc taggccctgc agacatgggt
     8401 gatcctcatg ttggcctagc tctaaactga ttggtccatg ccaccacggt gtttcgtcct
     8461 ttccacaaga tatataaagc caagaaatcg aaatactttc aagttacggt aagcatatga
     8521 tagtccattt taaaacataa ttttaaaact gcaaactacc caagaaatta ttactttcta
     8581 cgtcacgtat tttgtactaa tatctttgtg tttacagtca aattaattct aattatctct
     8641 ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcttcc
     8701 tgcccgacct tggcgatcca attctacc
//
