LOCUS       62056_6540        6724 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 6724)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6724
                     /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             2224..2250
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     polyA_signal    2291..2515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2561..2989
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3003..3332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3399..4190
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4367..4500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4537..4553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4561..4577)
                     /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(4585..4615)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4630..4651)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4939..5524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5698..6555)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6556..6660)
                     /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 cttggtaccg agctcggatc
      901 cactagtcca gtgtggtgga attcgccacc atgggggtat ctgattacat caaaggtctc
      961 ggtcgagcct tcggcacagg tttcactgac gcagtgtcta gggaagtgga agcgttgaag
     1021 aaccacttaa tcggctccga aggggctgtt gagaagatct tgaagaactt ggtgaagcta
     1081 atttcagcct tagttatagt catcagaagt gattatgata tggtcaccct cacagccaca
     1141 ctagctctga tcgggtgcca cgggagtcct tgggcgtgga tcaaatcaaa gacagcttcc
     1201 atactgggca ttcccatggc acaaaaacag agtgcctcat ggctaaagaa gttcaatgac
     1261 atggcaaatg ctgcaaaagg gcttgagtgg attttcaaca agatcagtaa gttcattgac
     1321 tggcttaaag agaagatcat tccagctgcc aaagagaaag ttgagttttt gaacaaccta
     1381 aaacagctcc ccttgttgga gaaccaagtc tccaatcttg aacagtctgc tgcctcacaa
     1441 gaagacttag aagctatgtt tggtaatgtg atatatctgg ctcacttttg ccgcaaattc
     1501 caaccactct acgcaactga ggccaagaga gtctacgctt tagagaaaag gatgaataac
     1561 tacatgcagt tcaagagcaa acaccgtatt gaacctgtat gcttgatcat cagaggttcc
     1621 cccggaacgg gcaaatcgct cgccacaggc attatagcta gagccattgc tgacaagtat
     1681 cgctctagtg tatactcact ccccccagac ccagatcact ttgatgggta taagcaacag
     1741 gtggtcgcgg tcatggatga tctctgccag aacccggacg gaaaagacat gtccctattt
     1801 tgtcaaatgg tttctacagt agattttgta ccacccatgg catcactaga ggagaaagga
     1861 gtgtccttca cctctaagtt tgtcattgca tcgaccaatg ctagtaacat catagtcccc
     1921 acagtttcag attcagatgc aattcgcagg cgattctata tggactgcga tatagaagtg
     1981 acagattctt acaagacaga cctcggtcgg ctggacgcag gtagagctgc caagctttgt
     2041 acagaaaata acactgctaa ttttaagaga tgcagcccac tggtgtgtgg taaggctatt
     2101 cagctgagag acaggaagtc caaagtgaga tatagcgtcg acaccgtggt atcggaactg
     2161 atcagagagt acaacaatag atctgctatt gggaatacta tagaagcact ctttcaactc
     2221 gagtacccat acgacgtacc agattacgct tagtctagag ggcccgttta aacccgctga
     2281 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2341 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2401 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2461 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2521 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2581 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2641 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2701 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2761 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2821 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2881 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2941 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3001 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3061 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3121 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3181 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3241 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3301 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     3361 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3421 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3481 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3541 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     3601 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3661 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3721 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3781 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3841 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3901 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3961 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4021 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4081 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4141 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4201 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4261 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4321 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4381 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4441 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4501 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5461 caccgctggt agcggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5521 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5581 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5641 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5701 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5761 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5821 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5881 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5941 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     6001 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6061 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6121 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6181 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6241 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6301 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6361 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6421 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6481 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6541 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6601 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6661 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     6721 tggt
//
