LOCUS       62056_16870        7583 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 7583)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(1664..3313)
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIVPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     promoter        complement(3338..3541)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(3542..3845)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        complement(3967..3985)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3997..4015)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4035..4051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4059..4075)
                     /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(4083..4113)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4128..4149)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4437..5025)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5199..6056)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6057..6161)
                     /label=AmpR promoter
     polyA_signal    complement(6249..6330)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(6563..7159)
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(7256..7572)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctagcggcc gcatacaaaa
      421 aaccaacaca cagatccaat gaaaataaaa gatcctttat taagcttaac tgtttaaacc
      481 tatgatatca cgcacgcgta acctcagcaa ctaaaaataa aaacttcagt atataacttg
      541 ggaacaaaaa agtagtaaag ggaagcaatt caagatctgt tattggtatt tgaatacatt
      601 tattgtgaca gaatgctgtt ataaatattc ataagacaaa ggccatcttt ttatctagga
      661 attgtcaaag agaagattcc aaattggaag gatacatctt ttgtaaaatc tgccaccaat
      721 tcctgctttg agaataagca cctattgtaa aatttctaac attataaatg gtcacagcac
      781 atgccacttg atgcaatcca aactttgaaa tgtttgactt ctcagtgggc tgtccctctc
      841 cactgcaacc ccccttcctc cagcctcctg aaacatcgca ctagcctttg gtaagcaatt
      901 ccatatagat acgctggggg agcgtggagt atgaagcctg acatagcatc aggtacatca
      961 ggtacattta tttctaagtc taatagagaa cagtttttac tgcttaatag taagaagcac
     1021 tgagagtgat tttatcgagc attcttaact ctttcaactc cacctagata gactaggatc
     1081 tacagatgat gcagaaaaga ggttagggga gtactttcca atagtttatt gtattttctt
     1141 aaatatcctt tctggaattt tcagaaacaa aatcatctgt aggctcagcc tcgctctcac
     1201 aaacctccct cctgccgccc ctccccaacc ctccccccgc tttgtgttct taaggagtac
     1261 tacagaagca atctacagtc tctattgcag tttgtaaccc cctccccctc ccccctttaa
     1321 tactgaatga gatcgaatgt taggtccatg cagttcttgg tcaatgttaa cgaaagtcca
     1381 acgttccgtt cgcgcgggga cagcccgtcc gcaaagcggg gcagcccgca ggcggccgct
     1441 ccctggctcc acgccaaggg agggcgcgac caagtccttc ccactcgtgc acactggggg
     1501 cgccgtcaga ccgaacccag tccaacttgg atacccttgg ctttagttct cggacacttc
     1561 ttttttctct ccgtcgcaac ttgtcaagtt ctcaagtctg tctctctgtg ttattttttt
     1621 tcttcgttgc ccctcagccc ccgacggtct ctcttcactc gagcaatttg gactttccgc
     1681 ccttcttggc ctttatgagg atctctctga tttttcttgc gtcgagtttt ccggtaagac
     1741 ctttcggtac ttcgtccaca aacacaactc ctccgcgcaa ctttttcgcg gttgttactt
     1801 gactggcgac gtaatccacg atctcttttt ccgtcatcgt ctttccgtgc tccaaaacaa
     1861 caacggcggc gggaagttca ccggcgtcat cgtcgggaag acctgccacg cccgcgtcga
     1921 agatgttggg gtgttgtaac aatatcgatt ccaattcagc gggggccacc tgatatcctt
     1981 tgtatttaat taaagacttc aagcggtcaa ctatgaagaa gtgttcgtct tcgtcccagt
     2041 aagctatgtc tccagaatgt agccatccat ccttgtcaat caaggcgttg gtcgcttccg
     2101 gattgtttac ataaccggac ataatcatag gtcctctgac acataattcg cctctctgat
     2161 taacgcccag cgttttcccg gtatccagat ccacaacctt cgcttcaaaa aatggaacaa
     2221 ctttaccgac cgcgcccggt ttatcatccc cctcgggtgt aatcagaata gctgatgtag
     2281 tctcagtgag cccatatcct tgtcgtatcc ctggaagatg gaagcgtttt gcaaccgctt
     2341 ccccgacttc tttcgaaaga ggtgcgcccc cagaagcaat ttcgtgtaaa ttagataaat
     2401 cgtatttgtc aatcagagtg cttttggcga agaatgaaaa tagggttggt actagcaacg
     2461 cactttgaat tttgtaatcc tgaagggatc gtaaaaacag ctcttcttca aatctataca
     2521 ttaagacgac tcgaaatcca catatcaaat atccgagtgt agtaaacatt ccaaaaccgt
     2581 gatggaatgg gacaacactt aaaatcgcag tatccggaac gatttgattg ccaaaaatag
     2641 gatctctggc atgcgagaat ctgacgcagg cagttctatg cggaagggcc acacccttag
     2701 gtaacccagt agatccagag gaattcatta tcagtgcaat tgttttgtca cgatcaaagg
     2761 actctggtac aaaatcgtat tcattaaaac cgggaggtag atgagatgtg acgaacgtgt
     2821 acatcgactg aaatccctgg taatccgttt tagaatccat gataataatt ttctggatta
     2881 ttggtaattt tttttgcacg ttcaaaattt tttgcaaccc ctttttggaa acaaacacta
     2941 cggtaggctg cgaaatgttc atactgttga gcaattcacg ttcattataa atgtcgttcg
     3001 cgggcgcaac tgcaactccg ataaataacg cgcccaacac cggcataaag aattgaagag
     3061 agttttcact gcatacgacg attctgtgat ttgtattcag cccatatcgt ttcatagctt
     3121 ctgccaaccg aacggacatt tcgaagtatt ccgcgtacgt gatgttcacc tcgatatgtg
     3181 catctgtaaa agcaattgtt ccaggaacca gggcgtatct cttcatagcc ttatgcagtt
     3241 gctctccagc ggttccatcc tctagaggat agaatggcgc cgggcctttc tttatgtttt
     3301 tggcgtcttc catggtggat ccacggttca ctaaaccagc tctgcttata tagacctccc
     3361 accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga cattttggaa
     3421 agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg tggagacttg
     3481 gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa aaccgcatca
     3541 ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag tcccataagg
     3601 tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca atagggggcg
     3661 tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt aaatactcca
     3721 cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac gtcattattg
     3781 acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc gtaagttatg
     3841 taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat tactattaat
     3901 aactaagatc tggtaccttg aattcatgct tctcctccct ttagtgaggg taattctctc
     3961 tctctcccta tagtgagtcg tattaattcc ttctcttcta tagtgtcacc taaatcgttg
     4021 caattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4081 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4141 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4201 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4261 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4321 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4381 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4441 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4501 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4561 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4621 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4681 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4741 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4801 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4861 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4921 ggaaaaagag ttggtagctc ttgatccggc aaaaaaacca ccgctggtag cggtggtttt
     4981 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5041 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5101 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5161 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5221 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5281 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5341 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5401 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5461 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5521 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5581 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5641 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5701 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5761 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5821 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5881 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5941 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6001 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6061 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6121 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6181 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     6241 acgagattgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     6301 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     6361 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     6421 atggctgatt atgatcctct agagtcggtg ggcctcgggg gcgggtgcgg ggtcggcggg
     6481 gccgccccgg gtggcttcgg tcggagccat ggggtcgtgc gctcctttcg gtcgggcgct
     6541 gcgggtcgtg gggcgggcgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     6601 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt
     6661 tgcggggcgc ggaggtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc
     6721 cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg
     6781 tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt
     6841 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
     6901 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
     6961 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
     7021 tgacccgctc gatgtggcgg tccgggtcga cggtgtggcg cgtggcgggg tagtcggcga
     7081 acgcggcggc gagggtgcgt acggcccggg ggacgtcgtc gcgggtggcg aggcgcaccg
     7141 tgggcttgta ctcggtcatg gaaggtcgtc tccttgtgag gggtcagggg cgtgggtcag
     7201 gggatggtgg cggcaccggt cgtggcggcc gacctgcagg catgcaagct agctttttgc
     7261 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggccgagg
     7321 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     7381 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     7441 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     7501 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     7561 actttccaca ccaatctttc gtc
//
