LOCUS       V021726                 6848 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021726
VERSION     V021726
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             941..2338
                     /label="ADRA1A(NM_000680)"
                     /note="ADRA1A(NM_000680)"
                     /gene="ADRA1A"
     CDS             2345..2410
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    2448..2672
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2718..3146
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3160..3489
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3556..4350
                     /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    4524..4645
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4694..4710)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4718..4734
                     /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(4742..4772)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4787..4808
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5096..5681)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5852..6712)
                     /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(6713..6817)
                     /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 atccgccacc atggtgtttc tctcgggaaa
      961 tgcttccgac agctccaact gcacccaacc gccggcaccg gtgaacattt ccaaggccat
     1021 tctgctcggg gtgatcttgg ggggcctcat tcttttcggg gtgctgggta acatcctagt
     1081 gatcctctcc gtagcctgtc accgacacct gcactcagtc acgcactact acatcgtcaa
     1141 cctggcggtg gccgacctcc tgctcacctc cacggtgctg cccttctccg ccatcttcga
     1201 ggtcctaggc tactgggcct tcggcagggt cttctgcaac atctgggcgg cagtggatgt
     1261 gctgtgctgc accgcgtcca tcatgggcct ctgcatcatc tccatcgacc gctacatcgg
     1321 cgtgagctac ccgctgcgct acccaaccat cgtcacccag aggaggggtc tcatggctct
     1381 gctctgcgtc tgggcactct ccctggtcat atccattgga cccctgttcg gctggaggca
     1441 gccggccccc gaggacgaga ccatctgcca gatcaacgag gagccgggct acgtgctctt
     1501 ctcagcgctg ggctccttct acctgcctct ggccatcatc ctggtcatgt actgccgcgt
     1561 ctacgtggtg gccaagaggg agagccgggg cctcaagtct ggcctcaaga ccgacaagtc
     1621 ggactcggag caagtgacgc tccgcatcca tcggaaaaac gccccggcag gaggcagcgg
     1681 gatggccagc gccaagacca agacgcactt ctcagtgagg ctcctcaagt tctcccggga
     1741 gaagaaagcg gccaaaacgc tgggcatcgt ggtcggctgc ttcgtcctct gctggctgcc
     1801 ttttttctta gtcatgccca ttgggtcttt cttccctgat ttcaagccct ctgaaacagt
     1861 ttttaaaata gtattttggc tcggatatct aaacagctgc atcaacccca tcatataccc
     1921 atgctccagc caagagttca aaaaggcctt tcagaatgtc ttgagaatcc agtgtctctg
     1981 cagaaagcag tcttccaaac atgccctggg ctacaccctg cacccgccca gccaggccgt
     2041 ggaagggcaa cacaaggaca tggtgcgcat ccccgtggga tcaagagaga ccttctacag
     2101 gatctccaag acggatggcg tttgtgaatg gaaatttttc tcttccatgc cccgtggatc
     2161 tgccaggatt acagtgtcca aagaccaatc ctcctgtacc acagcccggg tgagaagtaa
     2221 aagctttttg caggtctgct gctgtgtagg gccctcaacc cccagccttg acaagaacca
     2281 tcaagttcca accattaagg tccacaccat ctccctcagt gagaacgggg aggaagtcct
     2341 cgaggactac aaagaccatg acggtgatta taaagatcat gacatcgact acaaggatga
     2401 cgatgacaag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2461 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2521 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2581 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2641 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2701 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2761 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2821 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2881 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     2941 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3001 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3061 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3121 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3181 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3241 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3301 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3361 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3421 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3481 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3541 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3601 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3661 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3721 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3781 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3841 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3901 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3961 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4021 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4081 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4141 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4201 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4261 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4321 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4381 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4441 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4501 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4561 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4621 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4681 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4741 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4801 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4861 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4921 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4981 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5041 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5101 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5161 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5221 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5281 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5341 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5401 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5461 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5521 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5581 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     5641 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5701 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5761 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5821 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5881 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5941 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6001 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6061 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6121 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6181 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6241 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6301 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6361 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6421 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6481 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6541 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6601 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6661 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6721 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6781 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6841 ctgacgtc
//
