LOCUS       V044951                 8571 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044951
VERSION     V044951
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             776..2497
                     /label="SLC1A2(NM_004171)"
                     /note="SLC1A2(NM_004171)"
                     /gene="SLC1A2"
     CDS             2513..2569
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /transl_table=1
                     /translation="ATNFSLLKQAGDVEENPGP"
                     /label="P2A"
     CDS             2570..3280
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     misc_feature    3347..3935
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4001..4049
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4251..4679
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4693..5022
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5089..5883
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    6059..6180
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6229..6245)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6253..6269
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6277..6307)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6322..6343
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6631..7219)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7390..8250)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8251..8355)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
      661 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgg
      721 actctagagc tagcgtttaa acttaagctt ggtaccgagc tcggatcccg ccaccatggc
      781 atctacggaa ggtgccaaca atatgcccaa gcaggtggaa gtgcgaatgc acgacagtca
      841 tcttggctca gaggaaccca agcaccggca cctgggcctg cgcctgtgtg acaagctggg
      901 gaagaatctg ctgctcaccc tgacggtgtt tggtgtcatc ctgggagcag tgtgtggagg
      961 gcttcttcgc ttggcatctc ccatccaccc tgatgtggtt atgttaatag ccttcccagg
     1021 ggatatactc atgaggatgc taaaaatgct cattctccct ctaatcatct ccagcttaat
     1081 cacagggttg tcaggcctgg atgctaaggc tagtggccgc ttgggcacga gagccatggt
     1141 gtattacatg tccacgacca tcattgctgc agtactgggg gtcattctgg tcttggctat
     1201 ccatccaggc aatcccaagc tcaagaagca gctggggcct gggaagaaga atgatgaagt
     1261 gtccagcctg gatgccttcc tggaccttat tcgaaatctc ttccctgaaa accttgtcca
     1321 agcctgcttt caacagattc aaacagtgac gaagaaagtc ctggttgcac caccgccgga
     1381 cgaggaggcc aacgcaacca gcgctgttgt ctctctgttg aacgagactg tgactgaggt
     1441 gccggaggag actaagatgg ttatcaagaa gggcctggag ttcaaggatg ggatgaacgt
     1501 cttaggtctg atagggtttt tcattgcttt tggcatcgct atggggaaga tgggagatca
     1561 ggccaagctg atggtggatt tcttcaacat tttgaatgag attgtaatga agttagtgat
     1621 catgatcatg tggtactctc ccctgggtat cgcctgcctg atctgtggaa agatcattgc
     1681 aatcaaggac ttagaagtgg ttgctaggca actggggatg tacatggtaa cagtgatcat
     1741 aggcctcatc atccacgggg gcatctttct ccccttgatt tactttgtag tgaccaggaa
     1801 aaaccccttc tccttttttg ctggcatttt ccaagcttgg atcactgccc tgggcaccgc
     1861 ttccagtgct ggaactttgc ctgtcacctt tcgttgcctg gaagaaaatc tggggattga
     1921 taagcgtgtg actagattcg tccttcctgt tggagcaacc attaacatgg atggtacagc
     1981 cctttatgaa gcggtagccg ccatctttat agcccaaatg aatggtgttg tcctggatgg
     2041 aggacagatt gtgactgtaa gcctcacagc caccctggca agcgtcggcg cggccagtat
     2101 ccccagtgcc gggctggtca ccatgctcct cattctgaca gccgtgggcc tgccaacaga
     2161 ggacatcagc ctgctggtgg ctgtggactg gctgctggac aggatgagaa cttcagtcaa
     2221 tgttgtgggt gactcttttg gggctgggat agtctatcac ctctccaagt ctgagctgga
     2281 taccattgac tcccagcatc gagtgcatga agatattgaa atgaccaaga ctcaatccat
     2341 ttatgatgac atgaagaacc acagggaaag caactctaat caatgtgtct atgctgcaca
     2401 caactctgtc atagtagatg aatgcaaggt aactctggca gccaatggaa agtcagccga
     2461 ctgcagtgtt gaggaagaac cttggaaacg tgagaaagaa ttcggaagcg gagctactaa
     2521 cttcagcctg ctgaagcagg ctggcgacgt ggaggagaac cctggaccta tggtgagcaa
     2581 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     2641 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     2701 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     2761 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     2821 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     2881 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     2941 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     3001 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     3061 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     3121 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     3181 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     3241 ccgccactcc accggcggca tggacgagct gtacaagtaa ctcgagtcta gagggcccct
     3301 taagggttcg atccctaccg gttagtaatg agtttgatat ctcgacaatc aacctctgga
     3361 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3421 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3481 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3541 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3601 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3661 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3721 ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct gtgttgccac
     3781 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3841 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3901 gacgagtcgg atctcccttt gggccgcctc cccgcctgga aacgggggag gctaactgaa
     3961 acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa aagacagaat
     4021 aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4081 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4141 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4201 aggccctgcc atagcagatc tgcgcagctg gggctctagg gggtatcccc acgcgccctg
     4261 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4321 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4381 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     4441 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4501 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4561 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4621 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4681 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4741 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4861 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4921 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     4981 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     5041 atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa
     5101 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     5161 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     5221 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     5281 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     5341 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     5401 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     5461 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     5521 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     5581 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     5641 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     5701 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     5761 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     5821 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     5881 tgagcgggac tctggggttc gcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5941 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     6001 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     6061 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     6121 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     6181 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     6241 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     6301 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     6361 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     6421 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     6481 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     6541 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     6601 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     6661 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6721 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6781 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag
     6841 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6901 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6961 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     7021 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     7081 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     7141 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     7201 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     7261 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     7321 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     7381 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     7441 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     7501 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     7561 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     7621 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     7681 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7741 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7801 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7861 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7921 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7981 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     8041 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     8101 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     8161 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     8221 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     8281 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     8341 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     8401 tctcccgatc ccctatggtc gactctcagt acaatctgct ctgatgccgc atagttaagc
     8461 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     8521 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag g
//
