LOCUS       62056_8870        8291 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 8291)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        50..1228
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1443..2159
                     /codon_start=1
                     /label=EGFP
                     /note="enhanced GFP"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2166..3056
                     /codon_start=1
                     /label=HaloTag(R)
                     /note="modified bacterial dehalogenase that forms covalent
                     bonds with chloroalkane derivatives"
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWREPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    3067..3655
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3721..3769
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3971..4399
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4413..4742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4809..5600
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5779..5912
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5949..5965)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5973..5989)
                     /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(5997..6027)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6042..6063)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6351..6939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7113..7970)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7971..8075)
                     /label=AmpR promoter
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg gctccggtgc
       61 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
      121 caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
      181 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
      241 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc
      301 ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta cttccacctg
      361 gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg agagttcgag
      421 gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg
      481 gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc
      541 tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc aagatagtct
      601 tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg cgggcggcga
      661 cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc
      721 gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc
      781 gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc
      841 ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga cgcggcgctc
      901 gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt cctcagccgt
      961 cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt agttctcgag
     1021 cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg agtttcccca
     1081 cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat tctccttgga
     1141 atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag tggttcaaag
     1201 tttttttctt ccatttcagg tgtcgtgagg aattagcttg gtactggcta gccaccatga
     1261 ggcttcggga gccgctcctg agcggcagcg ccgcgatgcc aggcgcgtcc ctacagcggg
     1321 cctgccgcct gctcgtggcc gtctgcgctc tgcaccttgg cgtcaccctc gtttactacc
     1381 tggctggccg cgacctgagc cgcctgcccc aactggtcgg agtctccaca ccgctgggat
     1441 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1501 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1561 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1621 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     1681 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1741 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1801 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1861 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1921 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1981 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     2041 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     2101 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaaga
     2161 agcttatggc cgagatcggc accggcttcc ccttcgaccc ccactacgtg gaggtgctgg
     2221 gcgagcgcat gcactacgtg gacgtgggcc cccgcgacgg cacccccgtg ctgttcctgc
     2281 acggcaaccc caccagcagc tacgtgtggc gcaacatcat cccccacgtg gcccccaccc
     2341 accgctgcat cgcccccgac ctgatcggca tgggcaagag cgacaagccc gacctgggct
     2401 acttcttcga cgaccacgtg cgcttcatgg acgccttcat cgaggccctg ggcctggagg
     2461 aggtggtgct ggtgatccac gactggggca gcgccctggg cttccactgg gccaagcgca
     2521 accccgagcg cgtgaagggc atcgccttca tggagttcat ccgccccatc cccacctggg
     2581 acgagtggcc cgagttcgcc cgcgagacct tccaggcctt ccgcaccacc gacgtgggcc
     2641 gcaagctgat catcgaccag aacgtgttca tcgagggcac cctgcccatg ggcgtggtgc
     2701 gccccctgac cgaggtggag atggaccact accgcgagcc cttcctgaac cccgtggacc
     2761 gcgagcccct gtggcgcgag cccaacgagc tgcccatcgc cggcgagccc gccaacatcg
     2821 tggccctggt ggaggagtac atggactggc tgcaccagag ccccgtgccc aagctgctgt
     2881 tctggggcac ccccggcgtg ctgatccccc ccgccgaggc cgcccgcctg gccaagagcc
     2941 tgcccaactg caaggccgtg gacatcggcc ccggcctgaa cctgctgcag gaggacaacc
     3001 ccgacctgat cggcagcgag atcgcccgct ggctgagcac cctggagatc agcggctgat
     3061 ctcgacaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     3121 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     3181 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     3241 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     3301 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     3361 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     3421 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     3481 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     3541 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     3601 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga
     3661 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     3721 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     3781 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     3841 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     3901 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     3961 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4021 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     4081 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcggggcat ccctttaggg
     4141 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     4201 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4261 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     4321 tttgatttat aagggatttt ggggatttcg gcctattggt taaaaaatga gctgatttaa
     4381 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     4441 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     4501 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4561 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4621 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4681 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4741 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4801 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4861 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4921 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4981 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     5041 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     5101 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     5161 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     5221 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     5281 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     5341 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5401 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5461 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5521 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5581 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     5641 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5701 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5761 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5821 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5881 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5941 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6001 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6061 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6121 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6181 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6241 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6301 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6361 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6421 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6481 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6541 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6601 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6661 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6721 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6781 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6841 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6901 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6961 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7021 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7081 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7141 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7201 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7261 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7321 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7381 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7441 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7501 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7561 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7621 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7681 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7741 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7801 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7861 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     7921 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     7981 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8041 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8101 gacgtcgacg gatcgggaga tctcccgatc ccctatggtc gactctcagt acaatctgct
     8161 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     8221 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     8281 atctgcttag g
//
