LOCUS       V031974                 6732 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031974
VERSION     V031974
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"
     CDS             706..1458
                     /label="FGF23(NM_020638)"
                     /note="FGF23(NM_020638)"
                     /gene="FGF23"
     CDS             1465..1488
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHHHH"
                     /label="8xHis"
     misc_feature    1508..2096
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2162..2210
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2412..2840
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2854..3183
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3250..4044
                     /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    4220..4341
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4390..4406)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4414..4430
                     /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(4438..4468)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4483..4504
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4792..5380)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5551..6411)
                     /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(6412..6516)
                     /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 gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatgtt gggggcccgc
      721 ctcaggctct gggtctgtgc cttgtgcagc gtctgcagca tgagcgtcct cagagcctat
      781 cccaatgcct ccccactgct cggctccagc tggggtggcc tgatccacct gtacacagcc
      841 acagccagga acagctacca cctgcagatc cacaagaatg gccatgtgga tggcgcaccc
      901 catcagacca tctacagtgc cctgatgatc agatcagagg atgctggctt tgtggtgatt
      961 acaggtgtga tgagcagaag atacctctgc atggatttca gaggcaacat ttttggatca
     1021 cactatttcg acccggagaa ctgcaggttc caacaccaga cgctggaaaa cgggtacgac
     1081 gtctaccact ctcctcagta tcacttcctg gtcagtctgg gccgggcgaa gagagccttc
     1141 ctgccaggca tgaacccacc cccgtactcc cagttcctgt cccggaggaa cgagatcccc
     1201 ctaattcact tcaacacccc cataccacgg cggcacaccc ggagcgccga ggacgactcg
     1261 gagcgggacc ccctgaacgt gctgaagccc cgggcccgga tgaccccggc cccggcctcc
     1321 tgttcacagg agctcccgag cgccgaggac aacagcccga tggccagtga cccattaggg
     1381 gtggtcaggg gcggtcgagt gaacacgcac gctgggggaa cgggcccgga aggctgccgc
     1441 cccttcgcca agttcatcct cgagcaccat caccatcacc atcaccatta gggcccgata
     1501 tctcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     1561 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     1621 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     1681 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     1741 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     1801 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     1861 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg
     1921 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     1981 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     2041 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg
     2101 aaacggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     2161 acggcaataa aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg
     2221 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     2281 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     2341 cgcagccaac gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag
     2401 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2461 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2521 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca tccctttagg
     2581 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2641 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2701 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2761 ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg agctgattta
     2821 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2881 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     2941 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3001 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3061 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3121 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3181 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3241 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3301 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3361 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3421 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3481 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3541 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3601 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3661 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3721 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3781 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3841 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3901 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     3961 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4021 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgcgaaatga ccgaccaagc
     4081 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4141 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4201 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4261 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4321 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4381 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     4441 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     4501 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     4561 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     4621 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     4681 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     4741 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4801 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4861 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4921 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4981 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5041 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5101 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5161 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5221 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5281 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5341 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5401 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5461 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     5521 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5581 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5641 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5701 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5761 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5821 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5881 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5941 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6001 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6061 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6121 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6181 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6241 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6301 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6361 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6421 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6481 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6541 tgacgtcgac ggatcgggag atctcccgat cccctatggt cgactctcag tacaatctgc
     6601 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     6661 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     6721 aatctgctta gg
//
