LOCUS       V032507                 7424 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032507
VERSION     V032507
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             902..2917
                     /label="SLC5A2(NM_003041)"
                     /note="SLC5A2(NM_003041)"
                     /gene="SLC5A2"
     CDS             2927..2944
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     polyA_signal    2991..3215
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3261..3689
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3703..4032
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4099..4893
                     /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    5067..5188
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5237..5253)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5261..5277
                     /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(5285..5315)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5330..5351
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5639..6224)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6395..7255)
                     /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(7256..7360)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 catggaggag cacacagagg caggctcggc accagagatg ggggcccaga aggccctgat
      961 tgacaatcct gctgacatcc tagtcattgc tgcatatttc ctgctggtca ttggcgttgg
     1021 cttgtggtcc atgtgcagaa ccaacagagg cactgtgggc ggctacttcc tggcaggacg
     1081 cagcatggtg tggtggccgg ttggggcctc tctcttcgcc agcaacatcg gcagtggcca
     1141 ctttgtgggc ctggcaggga ctggcgctgc aagtggcttg gctgttgctg gattcgagtg
     1201 gaatgcgctc ttcgtggtgc tgctactggg ctggctgttt gcacccgtgt acctgacagc
     1261 gggggtcatc acgatgccac agtacctgcg caagcgcttc ggcggccgcc gcatccgcct
     1321 ctacctgtct gtgctctccc ttttcctgta catcttcacc aagatctcag tggacatgtt
     1381 ctccggagct gtattcatcc agcaggctct gggctggaac atctatgcct ccgtcatcgc
     1441 gcttctgggc atcaccatga tttacacggt gacaggaggg ctggccgcgc tgatgtacac
     1501 ggacacggta cagaccttcg tcattctggg gggcgcctgc atcctcatgg gttacgcctt
     1561 ccacgaggtg ggcgggtatt cgggtctctt cgacaaatac ctgggagcag cgacttcgct
     1621 gacggtgtcc gaggatccag ccgtgggaaa catctccagc ttctgctatc gaccccggcc
     1681 cgactcctac cacctgctcc ggcaccccgt gaccggggat ctgccgtggc ccgcgctgct
     1741 cctcggactc acaatcgtct cgggctggta ctggtgcagc gaccaggtca tcgtgcagcg
     1801 ctgcctggcc gggaagagcc tgacccacat caaggcgggc tgcatcctgt gtgggtacct
     1861 gaagctgacg cccatgtttc tcatggtcat gccaggcatg atcagccgca ttctgtaccc
     1921 agacgaggtg gcgtgcgtgg tgcctgaggt gtgcaggcgc gtgtgcggca cggaggtggg
     1981 ctgctccaac atcgcctacc cgcggctcgt cgtgaagctc atgcccaacg gtctgcgcgg
     2041 actcatgctg gcggtcatgc tggccgcgct catgtcctcg ctggcctcca tcttcaacag
     2101 cagcagcacg ctcttcacca tggacatcta cacgcgcctg cggccacgcg ccggcgaccg
     2161 cgagctgctg ctggtgggac ggctctgggt ggtgttcatc gtggtagtgt cggtggcctg
     2221 gcttcccgtg gtgcaggcgg cacagggcgg gcagctcttc gattacatcc aggcagtctc
     2281 tagctacctg gcaccgcccg tgtccgccgt cttcgtgctg gcgctcttcg tgccgcgcgt
     2341 taatgagcag ggcgccttct ggggactcat cgggggcctg ctgatgggcc tggcacgcct
     2401 gattcccgag ttctccttcg gctcgggcag ctgtgtgcag ccctcggcgt gcccagcttt
     2461 cctctgcggc gtgcactacc tctacttcgc cattgtgctg ttcttctgct ctggcctcct
     2521 caccctcacg gtctccctgt gcaccgcgcc catccccaga aagcacctcc accgcctggt
     2581 cttcagtctc cggcatagca aggaggaacg ggaggacctg gatgctgatg agcagcaagg
     2641 ctcctcactc cctgtacaga atgggtgccc agagagtgcc atggagatga atgagcccca
     2701 ggccccggca ccaagcctct tccgccagtg cctgctctgg ttttgtggaa tgagcagagg
     2761 tggggtgggc agtcctccgc cccttaccca ggaggaggca gcggcagcag ccaggcggct
     2821 ggaggacatc agcgaggacc cgagctgggc ccgtgtggtc aacctcaatg ccctgctcat
     2881 gatggcagtg gccgtgttcc tctggggctt ctatgccgcg gccgcacatc accatcatca
     2941 ccattgactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3001 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3061 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3121 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3181 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3241 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3301 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3361 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3421 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3481 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3541 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3601 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3661 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3721 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3781 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3841 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3901 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3961 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     4021 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4081 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4141 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4201 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4261 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4321 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4381 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4441 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4501 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4561 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4621 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4681 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4741 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4801 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4861 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4921 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4981 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5041 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5101 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5161 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5221 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5281 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5341 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5401 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5461 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5521 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5581 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5641 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5701 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5761 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5821 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5881 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5941 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6001 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6061 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6121 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt
     6181 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6241 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6301 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6361 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6421 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6481 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6541 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6601 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6661 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6721 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6781 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6841 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6901 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6961 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     7021 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     7081 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     7141 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     7201 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     7261 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7321 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     7381 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggt
//
