LOCUS       Exported                8547 bp DNA     circular SYN 30-AUG-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8547)
  AUTHORS   Dimitri Conomos
  TITLE     Direct Submission
  JOURNAL   Exported Feb 1, 2022 from SnapGene 5.3.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8547
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             910..1608
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     CDS             1642..3771
                     /codon_start=1
                     /product="There are a few mutations (K132R, K141E, & K142E 
                     and G680S) in the nucleolin but they are not thought to 
                     affect function."
                     /label=hNucleolin
                     /translation="MVKLAKAGKNQGDPKKMAPPPKEVEEDSEDEEMSEDEEDDSSGEE
                     VVIPQKKGKKAAATSAKKVVVSPTKKVAVATPAKKAAVTPGKKAAATPAKKTVTPAKAV
                     TTPGKKGATPGKALVATPGKKGAAIPARGAKNGKNAEEEDSDEEEDDDSEEDEEDDEDE
                     DEDEDEIEPAAMKAAAAAPASEDEDDEDDEDDEDDDDDEEDDSEEEAMETTPAKGKKAA
                     KVVPVKAKNVAEDEDEEEDDEDEDDDDDEDDEDDDDEDDEEEEEEEEEEPVKEAPGKRK
                     KEMAKQKAAPEAKKQKVEGTEPTTAFNLFVGNLNFNKSAPELKTGISDVFAKNDLAVVD
                     VRIGMTRKFGYVDFESAEDLEKALELTGLKVFGNEIKLEKPKGKDSKKERDARTLLAKN
                     LPYKVTQDELKEVFEDAAEIRLVSKDGKSKGIAYIEFKTEADAEKTFEEKQGTEIDGRS
                     ISLYYTGEKGQNQDYRGGKNSTWSGESKTLVLSNLSYSATEETLQEVFEKATFIKVPQN
                     QNGKSKGYAFIEFASFEDAKEALNSCNKREIEGRAIRLELQGPRGSPNARSQPSKTLFV
                     KGLSEDTTEETLKESFDGSVRARIVTDRETGSSKGFGFVDFNSEEDAKAAKEAMEDGEI
                     DGNKVTLDWAKPKGEGGFGGRGGGRGGFGGRGGGRGGRGGFGGRSRGGFGGRGGFRGGR
                     GGGGDHKPQGKKTKFE"
     CDS             3283..3294
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3798..3928
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3977..4201
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4247..4675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4689..5018
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4869..5004
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5067..6092
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    6222..6343
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6392..6408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6416..6432
                     /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(6440..6470)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6485..6506
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(6794..7379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7550..8410)
                     /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(8411..8515)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gctaccggta tgagcgagct gattaaggag aacatgcaca tgaagctgta catggagggc
      961 accgtggaca accatcactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc
     1021 acccagacca tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc
     1081 ctggctacta gcttcctcta cggcagcaag accttcatca accacaccca gggcatcccc
     1141 gacttcttca agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa
     1201 gacgggggcg tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac
     1261 aacgtcaaga tcagaggggt gaacttcaca tccaacggcc ctgtgatgca gaagaaaaca
     1321 ctcggctggg aggccttcac cgagacgctg taccccgctg acggcggcct ggaaggcaga
     1381 aacgacatgg ccctgaagct cgtgggcggg agccatctga tcgcaaacat caagaccaca
     1441 tatagatcca agaaacccgc taagaacctc aagatgcctg gcgtctacta tgtggactac
     1501 agactggaaa gaatcaagga ggccaacaac gagacctacg tcgagcagca cgaggtggca
     1561 gtggccagat actgcgacct ccctagcaaa ctggggcaca agcttaataa gtactcagat
     1621 ctcgagctca agcttggtac catggtgaag ctcgcgaagg caggtaaaaa tcaaggtgac
     1681 cccaagaaaa tggctcctcc tccaaaggag gtagaagaag atagtgaaga tgaggaaatg
     1741 tcagaagatg aagaagatga tagcagtgga gaagaggtcg tcatacctca gaagaaaggc
     1801 aagaaggctg ctgcaacctc agcaaagaag gtggtcgttt ccccaacaaa aaaggttgca
     1861 gttgccacac cagccaagaa agcagctgtc actccaggca aaaaggcagc agcaacacct
     1921 gccaagaaga cagttacacc agccaaagca gttaccacac ctggcaagaa gggagccaca
     1981 ccaggcaaag cattggtagc aactcctggt aagaagggtg ctgccatccc agccaggggg
     2041 gcaaagaatg gcaagaatgc cgaggaggaa gacagtgatg aagaggagga tgatgacagt
     2101 gaggaggatg aggaggatga cgaggacgag gatgaggatg aagatgaaat tgaaccagca
     2161 gcgatgaaag cagcagctgc tgcccctgcc tcagaggatg aggacgatga ggatgacgaa
     2221 gatgatgagg atgacgatga cgatgaggaa gatgactctg aagaagaagc tatggagact
     2281 acaccagcca aaggaaagaa agctgcaaaa gttgttcctg tgaaagccaa gaacgtggct
     2341 gaggatgaag atgaagaaga ggatgatgag gacgaggatg acgacgacga cgaagatgat
     2401 gaagatgatg atgatgaaga tgatgaggag gaggaagaag aggaggagga agagcctgtc
     2461 aaagaagcac ctggaaaacg aaagaaggaa atggccaaac agaaagcagc tcctgaagcc
     2521 aagaaacaga aagtggaagg cacagaaccg actacggctt tcaatctctt tgttggaaac
     2581 ctaaacttta acaaatctgc tcctgaatta aaaactggta tcagcgatgt ttttgctaaa
     2641 aatgatcttg ctgttgtgga tgtcagaatt ggtatgacta ggaaatttgg ttatgtggat
     2701 tttgaatctg ctgaagacct ggagaaagcg ttggaactca ctggtttgaa agtctttggc
     2761 aatgaaatta aactagagaa accaaaagga aaagacagta agaaagagcg agatgcgaga
     2821 acacttttgg ctaaaaatct cccttacaaa gtcactcagg atgaattgaa agaagtgttt
     2881 gaagatgctg cggagatcag attagtcagc aaggatggga aaagtaaagg gattgcttat
     2941 attgaattta agacagaagc tgatgcagag aaaacctttg aagaaaagca gggaacagag
     3001 atcgatgggc gatctatttc cctgtactat actggagaga aaggtcaaaa tcaagactat
     3061 agaggtggaa agaatagcac ttggagtggt gaatcaaaaa ctctggtttt aagcaacctc
     3121 tcctacagtg caacagaaga aactcttcag gaagtatttg agaaagcaac ttttatcaaa
     3181 gtaccccaga accaaaatgg caaatctaaa gggtatgcat ttatagagtt tgcttcattc
     3241 gaagacgcta aagaagcttt aaattcctgt aataaaaggg aaattgaggg cagagcaatc
     3301 aggctggagt tgcaaggacc caggggatca cctaatgcca gaagccagcc atccaaaact
     3361 ctgtttgtca aaggcctgtc tgaggatacc actgaagaga cattaaagga gtcatttgac
     3421 ggctccgttc gggctaggat agttactgac cgggaaactg ggtcctccaa agggtttggt
     3481 tttgtagact tcaacagtga ggaggatgcc aaagctgcca aggaggccat ggaagacggt
     3541 gaaattgatg gaaataaagt taccttggac tgggccaaac ctaagggtga aggtggcttc
     3601 gggggtcgtg gtggaggcag aggcggcttt ggaggacgag gtggtggtag aggaggccga
     3661 ggaggatttg gtggcagaag ccggggaggc tttggagggc gaggaggctt ccgaggaggc
     3721 agaggaggag gaggtgacca caagccacaa ggaaagaaga cgaagtttga atagctcgag
     3781 catgcatcta gaggatcaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3841 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3901 aactcatcaa tgtatcttat catgtctgga tccctcgagt ctagagggcc cgtttaaacc
     3961 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4021 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4081 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4141 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4201 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4261 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4321 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4381 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4441 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4501 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4561 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4621 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4681 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4741 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4801 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4861 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4921 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4981 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5041 ccattttcgg atctgatcag cacgtgatga aaaagcctga actcaccgcg acgtctgtcg
     5101 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     5161 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     5221 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     5281 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     5341 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     5401 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     5461 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     5521 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     5581 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     5641 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     5701 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     5761 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     5821 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     5881 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     5941 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     6001 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     6061 gccccagcac tcgtccgagg gcaaaggaat agcacgtgct acgagatttc gattccaccg
     6121 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     6181 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     6241 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     6301 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     6361 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6421 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6481 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6541 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6601 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     7021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     7081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     7141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     7201 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     7261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     7321 ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7381 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7441 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7501 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7561 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7621 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7681 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7741 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7801 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7861 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7921 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7981 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8041 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8101 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8161 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     8221 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     8281 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8341 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8401 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     8461 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8521 ttccccgaaa agtgccacct gacgtca
//
