LOCUS       V019713                 6812 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019713
VERSION     V019713
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             683..1492
                     /label="Hoxa5(NM_010453)"
                     /note="Hoxa5(NM_010453)"
                     /gene="Hoxa5"
     CDS             1499..1564
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1588..2176
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2242..2290
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2492..2920
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        2934..3263
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3330..4124
                     /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    4300..4421
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4470..4486)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4494..4510
                     /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(4518..4548)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4563..4584
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4872..5460)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5631..6491)
                     /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(6492..6596)
                     /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 ggatccgcca ccatgagctc ttattttgta aactcatttt gcggtcgcta
      721 tccaaatggc ccggactacc agttgcataa ttatggagat catagttccg tgagcgaaca
      781 attcagggac tcggcgagca tgcactccgg caggtacggc tacggctaca atggcatgga
      841 tctcagcgtc ggccgttcgg gttccggcca ctttggctcc ggcgagcgcg cccgcagcta
      901 cgcggctggg gccagtgcgg cgcccgccga gcccaggtac agccagccgg ccacgtccac
      961 gcactcgcca ccgcccgacc cgctgccctg ctcagcggtg gccccctcgc ccggcagcga
     1021 cagccaccac ggcgggaaaa actccctggg caactccagc ggcgcctcgg ccaacgccgg
     1081 cagcacccac atcagcagca gagagggggt tggcacggcg tccgcagccg aggaggacgc
     1141 ccctgccagc agcgagcagg cgggcgccca gagcgagccg agcccggcgc cgcccgctca
     1201 gccccagatc tacccctgga tgcgcaagct gcacattagt cacgacaata taggtggccc
     1261 agaaggcaaa agggcccgga cggcctacac tcgctaccag accctggagc tggagaaaga
     1321 attccacttc aaccgctacc tgacccgccg aagaaggatc gaaatagctc atgccctttg
     1381 cctctccgag agacaaatta aaatctggtt ccaaaacagg aggatgaagt ggaaaaaaga
     1441 taataagctg aaaagcatga gtatggccgc ggcagggggg gctttccgcc ccctcgagga
     1501 ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg atgacgatga
     1561 caagtagtga gggcccgata tctcgacaat caacctctgg attacaaaat ttgtgaaaga
     1621 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     1681 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     1741 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     1801 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     1861 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     1921 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     1981 aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2041 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2101 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     2161 tgggccgcct ccccgcctgg aaacggggga ggctaactga aacacggaag gagacaatac
     2221 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     2281 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     2341 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     2401 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     2461 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2521 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2581 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2641 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2701 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2761 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2821 aaccctatct cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg
     2881 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2941 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     3001 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3061 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3121 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3181 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3241 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3301 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3361 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3421 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3481 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3541 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3601 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3661 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3721 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3781 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3841 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3901 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3961 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     4021 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     4081 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4141 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4201 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4261 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4321 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4381 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4441 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4501 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4561 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4621 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4681 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4741 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4801 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5041 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     5101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5281 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     5341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5581 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5641 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5701 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5761 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5821 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5881 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5941 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6001 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6061 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6121 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6181 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6241 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6301 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6361 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6421 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6481 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6541 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6601 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6661 cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6721 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     6781 tgaccgacaa ttgcatgaag aatctgctta gg
//
