LOCUS       V040231                 7564 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V040231
VERSION     V040231
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             964..3054
                     /label="Slc34a2(NM_011402)"
                     /note="Slc34a2(NM_011402)"
                     /gene="Slc34a2"
     CDS             3061..3126
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    3164..3388
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3434..3862
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3876..4205
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4272..5066
                     /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    5240..5361
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5410..5426)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5434..5450
                     /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(5458..5488)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5503..5524
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5812..6397)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6568..7428)
                     /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(7429..7533)
                     /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 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc
      961 accatggctc cttggcccga gttggaaaat gcccagccca accccggtaa attcatcgag
     1021 ggggcctctg gaccacaatc gagcatacct gccaaagaca aggaggccag caaaacaaat
     1081 gacaacggga ctcctgtagc caagactgaa ctgctaccct cctattccgc cctggttctc
     1141 atagaggagc atcccgaggg gacggacccc tgggacctgc ctgaactcca ggacactggg
     1201 atcaaatggt cagagagaga caccaaaggg aagacgctct gcattttcca gggggttggg
     1261 aagttcattc ttctcctggg gtttctctac ttgttcgtgt gctccctgga tgtactgagc
     1321 agcgccttcc agcttgttgg aggcaaagtg gccggacagt tcttcagcaa caactccatc
     1381 atgtctaatc ctgtggctgg actggtgata ggggttctgg tgacagtcat ggttcaaagc
     1441 tccagcacct cttcctccat catcgtcagc atggttgctt cttcattgct gacggttcgg
     1501 gctgccatcc ccatcatcat gggcgctaac attggaacct ccatcaccaa cacgatcgtg
     1561 gcgcttatgc aggcaggaga cagaaatgag ttcagaaggg catttgcagg agccactgtt
     1621 catgacttct tcaactggct ctctgtgttc gtgctcttgc ctctggaggc tgccacccat
     1681 tacctggaga tcctgaccaa ccttgtgtta gaaaccttca agttccagaa tggagaagat
     1741 gccccagaca ttctgaaagt catcacagac cccttcacaa agctcatcat ccagctggat
     1801 aaaaaggtca tccaacaaat cgcaatgggt gactcagcgg cccagaacaa gagcctgatc
     1861 aagatctggt gtaaaagtat aaccaatgtg actgagatga atgtcaccgt cccctcgact
     1921 gataactgca cctctccttc atattgctgg actgatggca tacagacctg gaccatccag
     1981 aacgttaccc aaaaggagaa cattgctaaa tgtcagcaca tcttcgtgaa cttcagtctc
     2041 ccagaccttg ctgtgggcat catcttgctt actgtctccc tggtggtcct ctgtggctgt
     2101 ctgatcatga tagtcaagct gttgggttct gtgctcaggg gacaagtcgc tactgtcatc
     2161 aagaagaccc tgaatactga tttccccttt ccctttgctt ggctgactgg ctacctggcc
     2221 atccttgtgg gggcaggcat gaccttcatc gtgcaaagca gctctgtgtt cacgtctgcc
     2281 atgactccac tcatcggtat tggggtgatc agcattgaga gggcatatcc actcacgttg
     2341 ggctccaaca ttggcaccac caccaccgcc attctggctg ctttagccag cccaggcaac
     2401 accttgagga gttctctcca gattgccctg tgccatttct tcttcaacat ctctggaata
     2461 ttgctttggt acccaatccc attcacccgt ctgcccatcc gtctggctaa aggcctaggc
     2521 aacatctctg ccaagtatcg ctggttcgca gtcttctatc tcatcttctt cttctttgtg
     2581 accccgctga cagtgtttgg cctgtctctg gcaggctggc cagtgctggt gggcgtgggc
     2641 gtacccatca tcctgctgct gctcctggtt ctgtgcctcc ggatgctgca gttccgctgc
     2701 cctcgcatct tgcccctgaa gctccgggac tggaatttcc tacccttgtg gatgcactct
     2761 ctgaaaccct gggacaacgt catctccctg gccaccacct gcttccagag gcgttgctgc
     2821 tgctgttgcc gcgtgtgctg ccgtgtctgc tgcatggtgt gcggctgcaa gtgctgtcgc
     2881 tgcagcaagt gctgcaggga ccagggagag gaagaggagg agaaggagca ggatatcccg
     2941 gtcaaggctt ctggggcctt tgacaatgca gctatgagca aagagtgcca agatgagggc
     3001 aagggccaag tggaagttct gagcatgaag gccctgtcca acactacagt gttcctcgag
     3061 gactacaaag accatgacgg tgattataaa gatcatgaca tcgactacaa ggatgacgat
     3121 gacaagtagt gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3181 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3241 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3301 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3361 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3421 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3481 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3541 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3601 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3661 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3721 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3781 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3841 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3901 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3961 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4021 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4081 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4141 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4201 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4261 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4321 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4381 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4441 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4501 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4561 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4621 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4681 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4741 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4801 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4861 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4921 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4981 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5041 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     5101 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5161 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5221 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5341 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5401 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5461 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5521 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5581 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5641 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5701 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5761 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5821 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5881 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5941 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6001 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6061 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6121 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6181 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6241 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6301 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg
     6361 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6421 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6481 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6541 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6601 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6661 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6721 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6781 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6841 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6901 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6961 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     7021 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7081 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7141 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7201 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7261 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7321 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7381 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7441 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7501 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7561 cgtc
//
