LOCUS       V032696                 8345 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032696
VERSION     V032696
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             757..1980
                     /label="ZSCAN1(NM_182572)"
                     /note="ZSCAN1(NM_182572)"
                     /gene="ZSCAN1"
     CDS             2287..2343
                     /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             2344..3054
                     /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    3121..3709
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3775..3823
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4025..4453
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4467..4796
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4863..5657
                     /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    5833..5954
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6003..6019)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6027..6043
                     /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(6051..6081)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6096..6117
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6405..6993)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7164..8024)
                     /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(8025..8129)
                     /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 gccaccatgc ttccacggcc caaagcccct
      781 gcctccccca gacgccccca gaccccaacc ccgagtgagc aggacgcaga ccctgggcca
      841 gcaagcccca gggacaccga agcccagcgt ctgcgcttcc ggcagttcca gtaccacgtg
      901 gcgagcgggc cgcacctcgc gctgggccag ctctggacgc tgtgccgcca gtggctgagg
      961 cccgaggcgc gctccaagga gcagatgctg gagctgctgg tgctggagca gttcctgggc
     1021 gcgctgccca gcaagatgcg gacctgggtg cagtcacagg gcccccgaag ctgcagggag
     1081 gccgccagcc tggtggagga cctcacacag atgtgccagc aggaagttct ggtatctctg
     1141 gactcggtcg aaccccagga ctggagtttc ggtgaggagg aagatgggaa gagtccaagg
     1201 tcccagaaag aaccatcgca ggcgtctgag ctgattctgg atgcagtggc agcagcccca
     1261 gcactccccg aggaaagtga gtggctggag actacccagc tccagcagag tctgcacacc
     1321 agggcggagg ccgaagcgcc ccgcgcccct ggcttgctgg ggtcccgggc ccgcttgcct
     1381 ctgaagccga gtatctggga cgagcctgag gaccttctcg cagggccctc ctcagacctg
     1441 cgggcagaag ggactgtgat ctcgagcccc aagggtccaa gtgctcagag aatcagtccc
     1501 cgaaggagaa acaggaacac tgaccagagc ggccgccacc agccatccct caagcacacc
     1561 aaaggtggta cccaagaggc tgttgcaggc atctcggtag tgccgcgtgg gccccgaggt
     1621 gggcggccct tccagtgtgc cgactgtggg atggtcttca cctgggtcac ccacttcatc
     1681 gagcaccaga agacccatcg cgaggaaggg ccctttccgt gccccgagtg tggcaaggtc
     1741 ttcctgcaca actccgtcct cactgagcat ggcaagatcc acctgctgga gccaccgagg
     1801 aagaaagccc cccggagcaa gggcccccgg gagtccgtcc cacccaggga tggagcccag
     1861 ggcccagtgg cccctcgcag ccccaaaaga cccttccagt gtagcgtctg cgggaaggcc
     1921 ttcccctgga tggtccacct cattgaccac cagaagctcc acacggccca cggccacatg
     1981 gaattcgcca ccatgattct ctgggaaaag aaaagcgaag atattaccga gacattcaac
     2041 cccgacaaca acatgatcgc cgccgacgcg agcgaggcat cggaggcggg gaaggcagaa
     2101 aacacccatt acaccggcaa gcaccgcttc atgtaccgga tcctgttcgt cctgacctgc
     2161 ctgatcacct tcattgtggt ggctattgtc ggcggaatta tcgcttcgaa cagaaacgtc
     2221 ggaaatagaa tagctcaagg ccaaggatta gccttgaaat tagcaggtct tgatatcgga
     2281 agcggagcta ctaacttcag cctgctgaag caggctggcg acgtggagga gaaccctgga
     2341 cctatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
     2401 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
     2461 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
     2521 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
     2581 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
     2641 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
     2701 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
     2761 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
     2821 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
     2881 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
     2941 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
     3001 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaactcgag
     3061 tctagagggc cccttaaggg ttcgatccct accggttagt aatgagtttg atatctcgac
     3121 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     3181 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     3241 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     3301 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     3361 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     3421 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3481 ttgggcactg acaattccgt ggtgttgtcg gggaagctga cgtcctttcc atggctgctc
     3541 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3601 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3661 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggaaacggg
     3721 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3781 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3841 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3901 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3961 aacgtcgggg cggcaggccc tgccatagca gatctgcgca gctggggctc tagggggtat
     4021 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     4081 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     4141 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg gcatcccttt agggttccga
     4201 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     4261 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     4321 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     4381 ttataaggga ttttggggat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     4441 tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     4501 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     4561 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4621 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     4681 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct
     4741 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     4801 tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc
     4861 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4921 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4981 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     5041 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     5101 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     5161 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     5221 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     5281 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     5341 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     5401 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     5461 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     5521 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     5581 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5641 ttgacgagtt cttctgagcg ggactctggg gttcgcgaaa tgaccgacca agcgacgccc
     5701 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5761 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5821 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5881 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5941 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6001 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6061 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6121 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6181 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6241 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6301 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6361 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6421 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6481 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6541 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     6601 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6661 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6721 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6781 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6841 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6901 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6961 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7021 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7081 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7141 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7201 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7261 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7321 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7381 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7441 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7501 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7561 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7621 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7681 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7741 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7801 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7861 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7921 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7981 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8041 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8101 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8161 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
     8221 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
     8281 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
     8341 ttagg
//
