LOCUS       V023706                 6419 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023706
VERSION     V023706
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             779..1099
                     /label="HMGA1(NM_145899)"
                     /note="HMGA1(NM_145899)"
                     /gene="HMGA1"
     CDS             1106..1171
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1195..1783
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    1849..1897
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2099..2527
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2541..2870
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             2937..3731
                     /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    3907..4028
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4077..4093)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4101..4117
                     /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(4125..4155)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4170..4191
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4479..5067)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5238..6098)
                     /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(6099..6203)
                     /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 ctcggatcca ctagtccagt gtggtggaat tcgccaccat
      781 gagtgagtcg agctcgaagt ccagccagcc cttggcctcc aagcaggaaa aggacggcac
      841 tgagaagcgg ggccggggca ggccgcgcaa gcagcctccg gtgagtcccg ggacagcgct
      901 ggtagggagt cagaaggagc ccagcgaagt gccaacacct aagagacctc ggggccgacc
      961 aaagggaagc aaaaacaagg gtgctgccaa gacccggaaa accaccacaa ctccaggaag
     1021 gaaaccaagg ggcagaccca aaaaactgga gaaggaggaa gaggagggca tctcgcagga
     1081 gtcctcggag gaggagcagc tcgaggacta caaagaccat gacggtgatt ataaagatca
     1141 tgacatcgac tacaaggatg acgatgacaa gtagtgaggg cccgatatct cgacaatcaa
     1201 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     1261 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     1321 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     1381 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     1441 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     1501 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     1561 actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt
     1621 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     1681 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     1741 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcctggaaa cgggggaggc
     1801 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     1861 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     1921 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     1981 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     2041 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     2101 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2161 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2221 ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt ccgatttagt
     2281 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2341 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2401 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2461 gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2521 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2581 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2641 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2701 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2761 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2821 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2881 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2941 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3001 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3061 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3121 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3181 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3241 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3301 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3361 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3421 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3481 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3541 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3601 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3661 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3721 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     3781 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3841 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3901 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3961 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4021 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     4081 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     4141 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     4201 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     4261 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4321 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4381 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4441 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4501 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4561 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4621 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca
     4681 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4741 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4801 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4861 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     4921 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4981 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5041 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5101 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5161 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     5221 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     5281 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     5341 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     5401 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     5461 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5521 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5581 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5641 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5701 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5761 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5821 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5881 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5941 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6001 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6061 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6121 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6181 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     6241 tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct gatgccgcat
     6301 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     6361 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttagg
//
