LOCUS       62056_6885        6926 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6926)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6926
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        202..581
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        582..785
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        830..848
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2429..2446
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    2493..2717
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2763..3191
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3205..3534
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3601..4392
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4569..4702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4739..4755)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4763..4779)
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(4787..4817)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4832..4853)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5141..5726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5900..6757)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6758..6862)
                     /label=AmpR promoter
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttgccacca tgaagtgtgg
      901 catcgtccta gttgcgcttg ccttgttaat gctaggcagc tgtcccctca tccatgctct
      961 cttcagacag cctgcaataa tggatggaac actgcttgaa gttccggacc aggaagagga
     1021 ggagaggatg tgggtggacc caacccccaa aatcgaagat aacgatggca acgacgtgga
     1081 cgatgttggg accaaacgaa cgctggagga ggcagaggag cgagagaagg aagcaaagcg
     1141 aaaggctgac gaggaggagg aggagtttga acgactgatc catattgaaa aattcaaaag
     1201 gactttgttg caacgtctcc acttgacctc ccctcctgac ttcagccatc acggtggtat
     1261 ggccaaccga acacatggaa gacgtgtgct tcgatccctt ccattggctc ttcaaggacg
     1321 cctcttaaac cagatgcgtg ctgaagatgg aatggcagag ccaccaccgg atagaacgga
     1381 cgagagggaa actcttatcc ttctcaaaca cctgcactgg aggctaccaa aagttgcctc
     1441 tgctactttt ggaattgaaa tggctgatga catcgacccc tcgagaattc agtcggcttt
     1501 tctccgattt gaacacgttg aggtttggga gatcttcatg actccgagtg aggaggaagg
     1561 gaaaatgacg aacgcagcgg tagaccaacc gtctttggaa caatacaata acctgacgtg
     1621 gatgttcgaa cgacaaccca ccaaatacac gagtcttcca gcacccacag ttattcgtcg
     1681 atccttaggg tccccaatgg aacgccgtcg aactggttcc atccgcattc gtccgggtcg
     1741 tctggcggag acctttgtgc cgagttgtcc gggtttggtt caagtcacct ttgaaatcag
     1801 tggctccttc gcccagtgga tgtcacatag gcggcgaatg ccactgatgc gaaaactagt
     1861 ccgctctctc attgttgtct gtccggattg cagcagtcat gtagatcctg ttgatgttaa
     1921 caagggcatc cttgaaattc atcaccgcaa tatcgttcga cgaacacgga ggtcgcttga
     1981 caccaacagc tctcagcatg tccccatcgg gaatccgtgc agcccgaagg gacacaagtt
     2041 cagctgctgt acgcagccct tctcgttgaa cttagaggat gttggttgga ataactggat
     2101 tcttcatcca aaaaccgttg aacccaacta ctgtcacggt tcttgccaag ctgacgggat
     2161 tcagaagacc ccgcattctg acctgatgca cttgtaccgg agtcaaaatt acgaccgcct
     2221 ctcagaagtt cagcgggaag cgatgctctc ctgttgtcac cccgtcaaga tggcaagcac
     2281 cagtgtgctc tacgtggatc ccgacaatga gttgcacatg gacacccttc acaacatcat
     2341 tgtcctagag tgcggttgta gtggtaccga gctcggatcc actagtccag tgtggtggaa
     2401 ttctgcagat atcgccaccg cggccgcaca tcaccatcat caccattgac tcgagtctag
     2461 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
     2521 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
     2581 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
     2641 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
     2701 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
     2761 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2821 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2881 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2941 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3001 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3061 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3121 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3181 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     3241 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     3421 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
     3481 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
     3541 ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc
     3601 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3661 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3721 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3781 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3841 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3901 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3961 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4021 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4081 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4141 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4201 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4261 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4321 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4381 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4441 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4501 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4561 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4621 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     4681 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     4741 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     4801 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     4861 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     4921 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     4981 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     5041 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     5101 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5161 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5221 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5281 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5341 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5401 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5461 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5521 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5581 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     5641 gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga
     5701 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5761 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5821 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5881 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5941 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     6001 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     6061 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     6121 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     6181 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     6241 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     6301 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     6361 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6421 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6481 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6541 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6601 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6661 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6721 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6781 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6841 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     6901 cgggagatct cccgatcccc tatggt
//
