LOCUS       V031555                 9377 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031555
VERSION     V031555
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..3018
                     /label="SEMA3B(NM_001290061)"
                     /note="SEMA3B(NM_001290061)"
                     /gene="SEMA3B"
     CDS             3319..3375
                     /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             3376..4086
                     /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    4153..4741
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4807..4855
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      5057..5485
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        5499..5828
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5895..6689
                     /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    6865..6986
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7035..7051)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7059..7075
                     /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(7083..7113)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7128..7149
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7437..8025)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8196..9056)
                     /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(9057..9161)
                     /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 gccaccatgg ggcgggccgg ggctgccgcc
      781 gtgatcccgg gcctggccct gctctgggca gtggggctgg ggagtgccgc ccccagcccc
      841 ccacgccttc ggctctcctt ccaagagctc caggcctggc atggtctcca gactttcagc
      901 ctggagcgaa cctgctgcta ccaggccttg ctggtggatg aggagcgtgg acgcctgttt
      961 gtgggtgccg agaaccatgt ggcctccctc aacctggaca acatcagcaa gcgggccaag
     1021 aagctggcct ggccggcccc tgtggaatgg cgagaggagt gcaactgggc agggaaggac
     1081 attggtactg agtgcatgaa cttcgtgaag ttgctgcatg cctacaaccg cacccatttg
     1141 ctggcctgtg gcacgggagc cttccaccca acctgtgcct ttgtggaagt gggccaccgg
     1201 gcagaggagc ccgtcctccg gctggaccca ggaaggatag aggatggcaa ggggaagagt
     1261 ccttatgacc ccaggcatcg ggctgcctcc gtgctggtgg gggaggagct atactcaggg
     1321 gtggcagcag acctcatggg acgagacttt accatctttc gcagcctagg gcaacgtcca
     1381 agtctccgaa cagagccaca cgactcccgc tggctcaatg agcccaagtt tgtcaaggta
     1441 ttttggatcc cggagagcga gaacccagac gacgacaaaa tctacttctt ctttcgtgag
     1501 acggcggtag aggcggcgcc ggcactggga cgcctgtccg tgtcccgcgt tggccagatc
     1561 tgccggaacg acgtgggcgg ccagcgcagc ctggtcaaca agtggacgac gttcctgaag
     1621 gcgcggctgg tgtgctcggt gcccggcgtc gagggcgaca cccacttcga tcagctccgc
     1681 cccttccccg cagaggatgt gtttctgttg tcctcgcggg accaccggac cccgctgctc
     1741 tatgccgtct tctccacgtc cagcagcatc ttccagggct ctgcggtgtg cgtgtacagc
     1801 atgaacgacg tgcgccgggc cttcttggga ccctttgcac acaaggaggg gcccatgcac
     1861 cagtgggtgt cataccaggg tcgcgtcccc tacccgcggc caggcatgtg ccccagcaag
     1921 acctttggca ccttcagttc caccaaggac ttcccagacg atgtcatcca gtttgcgcgg
     1981 aaccaccccc tcatgtacaa ctctgtcctg cccactgggg ggcgccctct tttcctacaa
     2041 gttggagcca attacacctt cactcaaatt gccgcggacc gggttgcagc cgctgacgga
     2101 cactatgacg tcctcttcat tggcacagac gttggcacgg tgctgaaggt gatctcggtc
     2161 cccaagggca gtaggcccag cgcagagggg ctgctcctgg aggagctgca cgtgtttgag
     2221 gactcggccg ctgtcaccag catgcaaatt tcttccaaga ggcaccagct gtacgtagcc
     2281 tcgcggagcg cggtggccca gatcgcgttg caccgctgcg ctgcccacgg ccgcgtctgc
     2341 accgaatgct gtctggcgcg tgacccctac tgcgcctggg acggggtcgc gtgcacgcgc
     2401 ttccagccca gtgccaagag gcggttccgg cggcaagacg taaggaatgg cgaccccagc
     2461 acgttgtgct ccggagactc gtctcgtccc gcgctgctgg aacacaaggt gttcggcgtg
     2521 gagggcagca gcgcctttct ggagtgtgag ccccgctcgc tgcaggcgcg cgtggagtgg
     2581 actttccagc gcgcaggggt gacagcccac acccaggtgc tggcagagga gcgcaccgag
     2641 cgcaccgccc ggggactact gctgcgcagg ctgcggcgcc gggactcggg cgtgtacttg
     2701 tgcgccgccg tcgagcaggg ctttacgcaa ccgctgcgtc gcctgtcgct gcacgtgttg
     2761 agtgctacgc aggccgaacg actggcgcgg gccgaggagg ctgcgcccgc cgcgccgccg
     2821 ggccccaaac tctggtaccg ggactttctg cagctggtgg agccgggcgg aggtggcagc
     2881 gcgaactccc tgcgcatgtg ccgcccgcag cctgcgctgc agtcactgcc cctggagtcg
     2941 cggagaaagg gccgtaaccg gaggacccac gcccctgagc ctcgcgctga gcgggggccg
     3001 cgcagcgcaa cgcactggga attcatgatt ctctgggaaa agaaaagcga agatattacc
     3061 gagacattca accccgacaa caacatgatc gccgccgacg cgagcgaggc atcggaggcg
     3121 gggaaggcag aaaacaccca ttacaccggc aagcaccgct tcatgtaccg gatcctgttc
     3181 gtcctgacct gcctgatcac cttcattgtg gtggctattg tcggcggaat tatcgcttcg
     3241 aacagaaacg tcggaaatag aatagctcaa ggccaaggat tagccttgaa attagcaggt
     3301 cttgatatcg gaagcggagc tactaacttc agcctgctga agcaggctgg cgacgtggag
     3361 gagaaccctg gacctatggt gagcaagggc gaggaggata acatggccat catcaaggag
     3421 ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag
     3481 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
     3541 ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag
     3601 gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc
     3661 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
     3721 tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc
     3781 tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     3841 taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc
     3901 ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc
     3961 ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc
     4021 gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac
     4081 aagtaactcg agtctagagg gccccttaag ggttcgatcc ctaccggtta gtaatgagtt
     4141 tgatatctcg acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4201 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4261 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4321 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4381 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4441 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4501 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct gacgtccttt
     4561 ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4621 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4681 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4741 cctggaaacg ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4801 ctatgacggc aataaaaaga cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa
     4861 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4921 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4981 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cagatctgcg cagctggggc
     5041 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5101 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5161 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg gggcatccct
     5221 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     5281 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     5341 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     5401 tattcttttg atttataagg gattttgggg atttcggcct attggttaaa aaatgagctg
     5461 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     5521 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     5581 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5641 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     5701 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     5761 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     5821 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     5881 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5941 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6001 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6061 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6121 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6181 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6241 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6301 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6361 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6421 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6481 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6541 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6601 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6661 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     6721 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     6781 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     6841 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     6901 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     6961 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     7021 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7081 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7141 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7201 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7261 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7321 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7381 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7441 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7501 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     7561 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     7621 gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     7681 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     7741 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     7801 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     7861 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     7921 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     7981 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     8041 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     8101 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     8161 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     8221 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     8281 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     8341 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     8401 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     8461 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     8521 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     8581 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     8641 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     8701 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     8761 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     8821 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     8881 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     8941 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     9001 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     9061 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     9121 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     9181 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtcgact ctcagtacaa
     9241 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     9301 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     9361 tgaagaatct gcttagg
//
