LOCUS       62056_19440        6377 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 6377)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        192..495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        496..699
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             744..1457
                     /codon_start=1
                     /label=AcGFP1
                     /note="Aequorea coerulescens GFP"
                     /translation="MSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYFG
                     FVTAAAITHGMDELYK"
     polyA_signal    1475..1596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      1697..2125
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2139..2468
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2535..3326
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3503..3636
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3673..3689)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3697..3713)
                     /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(3721..3751)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3766..3787)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4075..4660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4834..5691)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5692..5796)
                     /label=AmpR promoter
     promoter        6062..6080
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        6211..6377
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
ORIGIN
        1 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg gtttatatat
       61 cttgtggaaa ggacgcggga tccgagctcg gtaccaagct taagtttaaa ccgctgatca
      121 gcctcgactg tgccttctaa atagtaatca attacggggt cattagttca tagcccatat
      181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      361 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      421 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      481 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      601 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      661 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      721 cgctagcgct accggtcgcc accatgagca agggcgccga gctgttcacc ggcatcgtgc
      781 ccatcctgat cgagctgaat ggcgatgtga atggccacaa gttcagcgtg agcggcgagg
      841 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      901 tgcctgtgcc ctggcccacc ctggtgacca ccctgagcta cggcgtgcag tgcttctcac
      961 gctaccccga tcacatgaag cagcacgact tcttcaagag cgccatgcct gagggctaca
     1021 tccaggagcg caccatcttc ttcgaggatg acggcaacta caagtcgcgc gccgaggtga
     1081 agttcgaggg cgataccctg gtgaatcgca tcgagctgac cggcaccgat ttcaaggagg
     1141 atggcaacat cctgggcaat aagatggagt acaactacaa cgcccacaat gtgtacatca
     1201 tgaccgacaa ggccaagaat ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     1261 atggcagcgt gcagctggcc gaccactacc agcagaatac ccccatcggc gatggccctg
     1321 tgctgctgcc cgataaccac tacctgtcca cccagagcgc cctgtccaag gaccccaacg
     1381 agaagcgcga tcacatgatc tacttcggct tcgtgaccgc cgccgccatc acccacggca
     1441 tggatgagct gtacaagtga ggactagata actgaacttg tttattgcag cttataatgg
     1501 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     1561 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtggga agacaatagc
     1621 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     1681 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     1741 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     1801 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     1861 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     1921 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     1981 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2041 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2101 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     2161 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2221 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2281 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2341 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2401 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2461 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     2521 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2581 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2641 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2701 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2761 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2821 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2881 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     2941 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3001 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3061 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3121 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     3181 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     3241 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     3301 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     3361 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     3421 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     3481 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag
     3541 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3601 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt
     3661 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     3721 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     3781 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     3841 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     3901 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     3961 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4021 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4081 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4141 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4201 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4261 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4321 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4381 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4441 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4501 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4561 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt
     4621 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4681 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4741 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4801 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4861 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4921 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4981 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5041 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5101 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5161 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5221 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5281 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5341 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5401 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5461 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5521 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5581 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5641 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5701 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5761 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5821 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     5881 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     5941 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     6001 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     6061 ttaatacgac tcactatagg gagagagaga gaattaccct cactaaaggg aggagaagca
     6121 tgaattcccc agtggaaaga cgcgcaggca aaacgcacca cgtgacggag cgtgaccgcg
     6181 cgccgagcgc gcgccaaggt cgggcaggaa gagggcctat ttcccatgat tccttcatat
     6241 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
     6301 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
     6361 ttaaaattat gttttaa
//
