LOCUS       62056_5525        8836 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 8836)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8836
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             673..1605
                     /codon_start=1
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             2420..4069
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     promoter        complement(4155..4173)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    4199..4423
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4469..4897
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4911..5240
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             5307..6098
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6275..6408
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6445..6461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6469..6485)
                     /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(6493..6523)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6538..6559)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6847..7435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7609..8466)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8467..8571)
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 gggctagcca ccatgacttc gaaagtttat gatccagaac aaaggaaacg gatgataact
      721 ggtccgcagt ggtgggccag atgtaaacaa atgaatgttc ttgattcatt tattaattat
      781 tatgattcag aaaaacatgc agaaaatgct gttatttttt tacatggtaa cgcggcctct
      841 tcttatttat ggcgacatgt tgtgccacat attgagccag tagcgcggtg tattatacca
      901 gaccttattg gtatgggcaa atcaggcaaa tctggtaatg gttcttatag gttacttgat
      961 cattacaaat atcttactgc atggtttgaa cttcttaatt taccaaagaa gatcattttt
     1021 gtcggccatg attggggtgc ttgtttggca tttcattata gctatgagca tcaagataag
     1081 atcaaagcaa tagttcacgc tgaaagtgta gtagatgtga ttgaatcatg ggatgaatgg
     1141 cctgatattg aagaagatat tgcgttgatc aaatctgaag aaggagaaaa aatggttttg
     1201 gagaataact tcttcgtgga aaccatgttg ccatcaaaaa tcatgagaaa gttagaacca
     1261 gaagaatttg cagcatatct tgaaccattc aaagagaaag gtgaagttcg tcgtccaaca
     1321 ttatcatggc ctcgtgaaat cccgttagta aaaggtggta aacctgacgt tgtacaaatt
     1381 gttaggaatt ataatgctta tctacgtgca agtgatgatt taccaaaaat gtttattgaa
     1441 tcggacccag gattcttttc caatgctatt gttgaaggtg ccaagaagtt tcctaatact
     1501 gaatttgtca aagtaaaagg tcttcatttt tcgcaagaag atgcacctga tgaaatggga
     1561 aaatatatca aatcgttcgt tgagcgagtt ctcaaaaatg aacaataatt ctagagcggc
     1621 ccccaagctt gggctgcagg tcttaaaaca gctctggggt tgtacccacc ccagaggccc
     1681 acgtggcggc tagtactccg gtattgcggt acctttgtac gcctgtttta tactcccttc
     1741 ccccgtaact tagaagcaca atgtccaagt tcaataggag ggggtacaaa ccagtaccac
     1801 cacgaacaag cacttctgtt cccccggtga ggctgtatag gctgtttcca cggctaaaag
     1861 cggctgatcc gttatccgct catgtacttc gagaagccta gtatcacctt ggaatcttcg
     1921 atgcgttgcg ctcaacactc aaccccagag tgtagcttag gtcgatgagt ctggacgttc
     1981 ctcaccggcg acggtggtcc aggctgcgtt ggcggcctac ctgtggccca aagccacagg
     2041 acgctagttg tgaacaaggt gtgaagagcc tattgagcta cctgagagtc ctccggcccc
     2101 tgaatgcggc taatcctaac cacggagcag gcagtggcaa tccagcgacc agcctgtcgt
     2161 aacgcgcaag ttcgtggcgg aaccgactac tttgggtgtc cgtgtttcct tttattttta
     2221 caatggctgc ttatggtgac aatcattgat tgttatcata aagcaaattg gattggccat
     2281 ccggtgagaa tttgattatt aaattactct cttgttggga ttgctccttt gaaatcttgt
     2341 gcactcacac ctattggaat tacctcattg ttaagatatc catcacactg gcggccgcat
     2401 ctagagggcc cggatccaaa tggaagacgc caaaaacata aagaaaggcc cggcgccatt
     2461 ctatcctcta gaggatggaa ccgctggaga gcaactgcat aaggctatga agagatacgc
     2521 cctggttcct ggaacaattg cttttacaga tgcacatatc gaggtgaaca tcacgtacgc
     2581 ggaatacttc gaaatgtccg ttcggttggc agaagctatg aaacgatatg ggctgaatac
     2641 aaatcacaga atcgtcgtat gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg
     2701 cgcgttattt atcggagttg cagttgcgcc cgcgaacgac atttataatg aacgtgaatt
     2761 gctcaacagt atgaacattt cgcagcctac cgtagtgttt gtttccaaaa aggggttgca
     2821 aaaaattttg aacgtgcaaa aaaaattacc aataatccag aaaattatta tcatggattc
     2881 taaaacggat taccagggat ttcagtcgat gtacacgttc gtcacatctc atctacctcc
     2941 cggttttaat gaatacgatt ttgtaccaga gtcctttgat cgtgacaaaa caattgcact
     3001 gataatgaat tcctctggat ctactgggtt acctaagggt gtggcccttc cgcatagaac
     3061 tgcctgcgtc agattctcgc atgccagaga tcctattttt ggcaatcaaa tcattccgga
     3121 tactgcgatt ttaagtgttg ttccattcca tcacggtttt ggaatgttta ctacactcgg
     3181 atatttgata tgtggatttc gagtcgtctt aatgtataga tttgaagaag agctgttttt
     3241 acgatccctt caggattaca aaattcaaag tgcgttgcta gtaccaaccc tattttcatt
     3301 cttcgccaaa agcactctga ttgacaaata cgatttatct aatttacacg aaattgcttc
     3361 tgggggcgca cctctttcga aagaagtcgg ggaagcggtt gcaaaacgct tccatcttcc
     3421 agggatacga caaggatatg ggctcactga gactacatca gctattctga ttacacccga
     3481 gggggatgat aaaccgggcg cggtcggtaa agttgttcca ttttttgaag cgaaggttgt
     3541 ggatctggat accgggaaaa cgctgggcgt taatcagaga ggcgaattat gtgtcagagg
     3601 acctatgatt atgtccggtt atgtaaacaa tccggaagcg accaacgcct tgattgacaa
     3661 ggatggatgg ctacattctg gagacatagc ttactgggac gaagacgaac acttcttcat
     3721 agttgaccgc ttgaagtctt taattaaata caaaggatat caggtggccc ccgctgaatt
     3781 ggaatcgata ttgttacaac accccaacat cttcgacgcg ggcgtggcag gtcttcccga
     3841 cgatgacgcc ggtgaacttc ccgccgccgt tgttgttttg gagcacggaa agacgatgac
     3901 ggaaaaagag atcgtggatt acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg
     3961 aggagttgtg tttgtggacg aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa
     4021 aatcagagag atcctcataa aggccaagaa gggcggaaag tccaaattgt aaaatgtaac
     4081 tgtattcagc gatgacgaaa ttcttagcta ttgtaatcct ccgaggcctc gagcatgcat
     4141 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     4201 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4261 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4321 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4381 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4441 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4621 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4801 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     4861 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4921 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4981 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5041 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5101 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5161 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5221 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     5281 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     5341 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     5401 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     5461 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     5521 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     5581 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     5641 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     5701 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5761 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5821 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5881 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5941 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6001 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6061 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     6121 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     6181 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     6241 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     6301 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     6361 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     6421 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     6481 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6541 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6601 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     6661 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6721 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6781 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     7321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8581 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     8641 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     8701 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     8761 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
     8821 gccagatata cgcgtt
//
