LOCUS       V019816                 6908 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019816
VERSION     V019816
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..2440
                     /label="SLC16A1(NM_003051)"
                     /note="SLC16A1(NM_003051)"
                     /gene="SLC16A1"
     CDS             2447..2470
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     polyA_signal    2508..2732
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      2778..3206
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3220..3549
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3616..4410
                     /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    4584..4705
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4754..4770)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4778..4794
                     /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(4802..4832)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4847..4868
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5156..5741)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5912..6772)
                     /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(6773..6877)
                     /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 atgccaccag cagttggagg
      961 tccagttgga tacacccccc cagatggagg ctggggctgg gcagtggtaa ttggagcttt
     1021 catttccatc ggcttctctt atgcatttcc caaatcaatt actgtcttct tcaaagagat
     1081 tgaaggtata ttccatgcca ccaccagcga agtgtcatgg atatcctcca taatgttggc
     1141 tgtcatgtat ggtggaggtc ctatcagcag tatcctggtg aataaatatg gaagtcgtat
     1201 agtcatgatt gttggtggct gcttgtcagg ctgtggcttg attgcagctt ctttctgtaa
     1261 caccgtacag caactatacg tctgtattgg agtcattgga ggtcttgggc ttgccttcaa
     1321 cttgaatcca gctctgacca tgattggcaa gtatttctac aagaggcgac cattggccaa
     1381 cggactggcc atggcaggca gccctgtgtt cctctgtact ctggcccccc tcaatcaggt
     1441 tttcttcggt atctttggat ggagaggaag ctttctaatt cttgggggct tgctactaaa
     1501 ctgctgtgtt gctggagccc tcatgcgacc aatcgggccc aagccaacca aggcagggaa
     1561 agataagtct aaagcatccc ttgagaaagc tggaaaatct ggtgtgaaaa aagatctgca
     1621 tgatgcaaat acagatctta ttggaagaca ccctaaacaa gagaaacgat cagtcttcca
     1681 aacaattaat cagttcctgg acttaaccct attcacccac agaggctttt tgctatacct
     1741 ctctggaaat gtgatcatgt tttttggact ctttgcacct ttggtgtttc ttagtagtta
     1801 tgggaagagt cagcattatt ctagtgagaa gtctgccttc cttctttcca ttctggcttt
     1861 tgttgacatg gtagcccgac catctatggg acttgtagcc aacacaaagc caataagacc
     1921 tcgaattcag tatttctttg cggcttccgt tgttgcaaat ggagtgtgtc atatgctagc
     1981 acctttatcc actacctatg ttggattctg tgtctatgcg ggattctttg gatttgcctt
     2041 cgggtggctc agctccgtat tgtttgaaac attgatggac cttgttggac cccagaggtt
     2101 ctccagcgct gtgggattgg tgaccattgt ggaatgctgt cctgtcctcc tggggccacc
     2161 acttttaggt cggctcaatg acatgtatgg agactacaaa tacacatact gggcatgtgg
     2221 cgtcgtccta attatttcag gtatctatct cttcattggc atgggcatca attatcgact
     2281 tttggcaaaa gaacagaaag caaacgagca gaaaaaggaa agtaaagagg aagagaccag
     2341 tatagatgtt gctgggaagc caaatgaagt taccaaagca gcagaatctc cggaccagaa
     2401 agacacagat ggagggccca aggaggagga aagtccagtc ctcgaggatt acaaggatga
     2461 cgacgataag tagtgagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag
     2521 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     2581 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     2641 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     2701 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     2761 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2821 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2881 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2941 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3001 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3061 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3121 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3181 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     3241 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3301 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3361 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     3421 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     3481 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     3541 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     3601 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     3661 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     3721 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     3781 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     3841 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     3901 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     3961 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     4021 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     4081 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     4141 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     4201 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     4261 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     4321 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     4381 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     4441 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4501 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4561 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4621 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4681 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4741 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4801 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4861 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4921 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     4981 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5041 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5101 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5161 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5221 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5281 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5341 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5401 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5461 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5521 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5581 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5641 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtttttttg
     5701 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5761 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5821 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5881 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5941 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6001 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6061 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6121 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6181 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6241 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6301 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6361 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6421 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6481 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6541 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6601 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6661 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6721 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6781 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6841 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6901 ctgacgtc
//
