LOCUS       V037199                 7932 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V037199
VERSION     V037199
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"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..2612
                     /label="SLC7A3(NM_032803)"
                     /note="SLC7A3(NM_032803)"
                     /gene="SLC7A3"
     CDS             2619..2684
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    2708..3296
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3362..3410
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3612..4040
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4054..4383
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4450..5244
                     /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    5420..5541
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5590..5606)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5614..5630
                     /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(5638..5668)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5683..5704
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5992..6580)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6751..7611)
                     /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(7612..7716)
                     /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 gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgcc gtggcaagca tttcgcagat
      781 ttggtcaaaa gctggtacgc agacgtacac tggagtcagg catggctgag actcgccttg
      841 ccagatgcct aagcaccctg gatttagtgg ccctgggtgt gggcagcaca ttgggtgcag
      901 gcgtgtatgt cctagctggc gaggtggcca aagataaagc agggccatcc attgtgatct
      961 gctttttggt ggctgccctg tcttctgtgt tggctgggct gtgctatgcg gagtttggtg
     1021 cccgggttcc ccgttctggt tcggcatatc tctacagcta tgtcactgtg ggtgaactct
     1081 gggccttcac cactggctgg aacctcatcc tctcctatgt cattggtaca gccagtgtgg
     1141 cccgggcctg gagctctgct tttgacaacc tgattgggaa ccacatctct aagactctgc
     1201 aggggtccat tgcactgcac gtgccccatg tccttgcaga atatccagat ttctttgctt
     1261 tgggcctcgt gttgctgctc actggattgt tggctctcgg ggctagtgag tcggccctgg
     1321 ttaccaaagt gttcacaggc gtgaaccttt tggttcttgg gttcgtcatg atctctggct
     1381 tcgttaaggg ggacgtgcac aactggaagc tcacagaaga ggactacgaa ttggccatgg
     1441 ctgaactcaa tgacacctat agcttgggtc ctctgggctc tggaggattt gtgcctttcg
     1501 gcttcgaggg aattctccgt ggagcagcga cctgtttcta tgcatttgtt ggtttcgact
     1561 gtattgctac cactggagaa gaagcccaga atccccagcg ttccatcccg atgggcattg
     1621 tgatctcact gtctgtctgc tttttggcgt attttgctgt ctcttctgca ctcaccctga
     1681 tgatgcctta ctaccagctt cagcctgaga gccctttgcc tgaggcattt ctctacattg
     1741 gatgggctcc tgcccgctat gttgtggctg ttggctccct ctgtgctctt tctaccagcc
     1801 tcctgggctc catgttcccc atgcctcggg tgatctacgc gatggcagag gatggcctcc
     1861 tgttccgtgt acttgctcgg atccacaccg gcacacgcac cccaatcata gccaccgtgg
     1921 tctctggcat tattgcagca ttcatggcat tcctcttcaa actcactgat cttgtggacc
     1981 tcatgtcaat tgggaccctg cttgcttact ccctggtgtc gatttgtgtt ctcatcctca
     2041 ggtatcaacc tgatcaggag acaaagactg gggaagaagt ggagttgcag gaggaggcaa
     2101 taactactga atcagagaag ttgaccctat ggggactatt tttcccactc aactccatcc
     2161 ccactccact ctctggccaa attgtctatg tttgttcctc attgcttgct gtcctgctga
     2221 ctgctctttg cctggtgctg gcccagtggt cagttccatt gctttctgga gacctgctgt
     2281 ggactgcagt ggttgtgctg ctcctgctgc tcattattgg gatcattgtg gtcatctgga
     2341 gacagccaca gagttccact ccccttcact ttaaggtgcc tgctttgcct ctcctcccac
     2401 taatgagcat ctttgtgaat atttacctta tgatgcagat gacagctggt acctgggccc
     2461 gatttggggt ctggatgctg attggctttg ctatctactt cggctatggg atccagcaca
     2521 gcctggaaga gattaagagt aaccaaccct cacgcaagtc tagagccaaa actgtagacc
     2581 ttgatcccgg cactctctat gtccactcag ttggatccga ctacaaagac catgacggtg
     2641 attataaaga tcatgacatc gactacaagg atgacgatga caagtagtga gggcccgata
     2701 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2761 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2821 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2881 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     2941 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3001 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3061 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     3121 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3181 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3241 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     3301 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     3361 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     3421 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     3481 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     3541 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     3601 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3661 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3721 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     3781 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3841 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3901 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3961 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     4021 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4081 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4141 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4201 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4261 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4321 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4381 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4441 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4501 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4561 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4621 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4681 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4741 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4801 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4861 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4921 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4981 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     5041 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5101 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5161 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5221 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     5281 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     5341 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     5401 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     5461 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     5521 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     5581 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5641 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5701 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5761 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5821 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5881 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5941 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6001 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6061 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6121 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6181 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6241 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6301 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6361 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6421 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6481 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6541 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6601 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6661 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6721 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6781 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6841 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6901 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6961 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7021 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7081 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7141 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7201 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7261 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7321 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7381 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7441 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7501 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     7561 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7681 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7741 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     7801 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7861 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     7921 aatctgctta gg
//
