LOCUS       pEnCMV-NLN(human)-HA    8135 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030585
VERSION     V030585
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             683..2794
                     /label="NLN(NM_020726)"
                     /note="NLN(NM_020726)"
                     /gene="NLN"
     CDS             2801..2827
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2831..2857
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2861..2887
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2911..3499
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3565..3613
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3815..4243
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4257..4586
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4653..5447
                     /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    5623..5744
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5793..5809)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5817..5833
                     /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(5841..5871)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5886..5907
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6195..6783)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6954..7814)
                     /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(7815..7919)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccgcca ccatgatcgc ccggtgcctt ttggctgtgc gaagcctccg
      721 cagagttggt ggttccagga ttttactcag aatgacgtta ggaagagaag tgatgtctcc
      781 tcttcaggca atgtcttcct atactgtggc tggcagaaat gttttaagat gggatctttc
      841 accagagcaa attaaaacaa gaactgagga gctcattgtg cagaccaaac aggtgtacga
      901 tgctgttgga atgctcggta ttgaggaagt aacttacgag aactgtctgc aggcactggc
      961 agatgtagaa gtaaagtata tagtggaaag gaccatgcta gactttcccc agcatgtatc
     1021 ctctgacaaa gaagtacgag cagcaagtac agaagcagac aaaagacttt ctcgttttga
     1081 tattgagatg agcatgagag gagatatatt tgagagaatt gttcatttac aggaaacctg
     1141 tgatctgggg aagataaaac ctgaggccag acgatacttg gaaaagtcaa ttaaaatggg
     1201 gaaaagaaat gggctccatc ttcctgaaca agtacagaat gaaatcaaat caatgaagaa
     1261 aagaatgagt gagctatgta ttgattttaa caaaaacctc aatgaggatg ataccttcct
     1321 tgtattttcc aaggctgaac ttggtgctct tcctgatgat ttcattgaca gtttagaaaa
     1381 gacagatgat gacaagtata aaattacctt aaaatatcca cactatttcc ctgtcatgaa
     1441 gaaatgttgt atccctgaaa ccagaagaag gatggaaatg gcttttaata caaggtgcaa
     1501 agaggaaaac accataattt tgcagcagct actcccactg cgaaccaagg tggccaaact
     1561 actcggttat agcacacatg ctgacttcgt ccttgaaatg aacactgcaa agagcacaag
     1621 ccgcgtaaca gcctttctag atgatttaag ccagaagtta aaacccttgg gtgaagcaga
     1681 acgagagttt attttgaatt tgaagaaaaa ggaatgcaaa gacaggggtt ttgaatatga
     1741 tgggaaaatc aatgcctggg atctatatta ctacatgact cagacagagg aactcaagta
     1801 ttccatagac caagagttcc tcaaggaata cttcccaatt gaggtggtca ctgaaggctt
     1861 gctgaacacc taccaggagt tgttgggact ttcatttgaa caaatgacag atgctcatgt
     1921 ttggaacaag agtgttacac tttatactgt gaaggataaa gctacaggag aagtattggg
     1981 acagttctat ttggacctct atccaaggga aggaaaatac aatcatgcgg cctgcttcgg
     2041 tctccagcct ggctgccttc tgcctgatgg aagccggatg atggcagtgg ctgccctcgt
     2101 ggtgaacttc tcacagccag tggcaggtcg tccctctctc ctgagacacg acgaggtgag
     2161 gacttacttt catgagtttg gtcacgtgat gcatcagatt tgtgcacaga ctgattttgc
     2221 acgatttagc ggaacaaatg tggaaactga ctttgtagag gtgccatcgc aaatgcttga
     2281 aaattgggtg tgggacgtcg attccctccg aagattgtca aaacattata aagatggaag
     2341 ccctattgca gacgatctgc ttgaaaaact tgttgcttct aggctggtca acacaggtct
     2401 tctgaccctg cgccagattg ttttgagcaa agttgatcag tctcttcata ccaacacatc
     2461 gctggatgct gcaagtgaat atgccaaata ctgctcagaa atattaggag ttgcagctac
     2521 tccaggcaca aatatgccag ctacctttgg acatttggca gggggatacg atggccaata
     2581 ttatggatat ctttggagtg aagtattttc catggatatg ttttacagct gttttaaaaa
     2641 agaagggata atgaatccag aggttggaat gaaatacaga aacctaatcc tgaaacctgg
     2701 gggatctctg gacggcatgg acatgctcca caatttcttg aaacgtgagc caaaccaaaa
     2761 agcgttccta atgagtagag gcctgcatgc tccgctcgag tacccatacg acgtacctga
     2821 ttatgcaact tatccttatg atgtacctga ttatgctagc tatccttatg atgtcccaga
     2881 ctacgcatag tgagggcccg atatctcgac aatcaacctc tggattacaa aatttgtgaa
     2941 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     3001 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     3061 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     3121 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     3181 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     3241 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     3301 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     3361 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     3421 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3481 ctttgggccg cctccccgcc tggaaacggg ggaggctaac tgaaacacgg aaggagacaa
     3541 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacgggtgtt
     3601 gggtcgtttg ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca
     3661 ccgagacccc attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca
     3721 agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagca
     3781 gatctgcgca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3841 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3901 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3961 ctaaatcggg gcatcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4021 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4081 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4141 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttggggat ttcggcctat
     4201 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt
     4261 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4321 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     4381 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     4441 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     4501 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     4561 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat
     4621 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4681 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4741 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4801 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc
     4861 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4921 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4981 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     5041 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5101 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5161 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5221 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5281 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5341 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5401 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5461 gttcgcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc
     5521 gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc
     5581 ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct
     5641 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5701 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg
     5761 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     5821 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     5881 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     5941 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     6001 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     6061 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     6121 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     6181 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     6241 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6301 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6361 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     6421 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6481 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6541 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6601 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     6661 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6721 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6781 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6841 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6901 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6961 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7021 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7081 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7141 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     7201 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     7261 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7321 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7381 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7441 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7501 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7561 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7621 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7681 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7741 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7801 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7861 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7921 acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat
     7981 ggtcgactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg
     8041 cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag
     8101 gcttgaccga caattgcatg aagaatctgc ttagg
//
