LOCUS       62056_20205        7235 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 7235)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        156..485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        533..580
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             599..994
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    1155..1288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1325..1341)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1349..1365)
                     /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(1373..1403)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1418..1439)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1727..2315)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2489..3346)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3347..3451)
                     /label=AmpR promoter
     enhancer        3717..4096
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4097..4300
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        4345..4363
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4392..4412
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             4422..4445
                     /codon_start=1
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
                     /translation="DYKDDDDK"
     CDS             5193..5219
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             5331..6017
                     /codon_start=1
                     /label=APOBEC-1
                     /note="cytidine deaminase (C to U editing enzyme) from rat"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     CDS             6033..6281
                     /codon_start=1
                     /label=UGI
                     /note="uracil-DNA glycosylase inhibitor from a Bacillus
                     subtilis bacteriophage (Mol et al., 1995)"
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             6339..6500
                     /codon_start=1
                     /label=GB1
                     /note="B1 domain of Streptococcal protein G"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             6528..6548
                     /codon_start=1
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /translation="PKKKRKV"
     CDS             6582..6623
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             6633..6650
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    6679..6903
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6949..7235
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
       61 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      121 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      421 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa
      541 ttaatcatcg gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat
      601 ggccaagcct ttgtctcaag aagaatccac cctcattgaa agagcaacgg ctacaatcaa
      661 cagcatcccc atctctgaag actacagcgt cgccagcgca gctctctcta gcgacggccg
      721 catcttcact ggtgtcaatg tatatcattt tactggggga ccttgtgcag aactcgtggt
      781 gctgggcact gctgctgctg cggcagctgg caacctgact tgtatcgtcg cgatcggaaa
      841 tgagaacagg ggcatcttga gcccctgcgg acggtgccga caggtgcttc tcgatctgca
      901 tcctgggatc aaagccatag tgaaggacag tgatggacag ccgacggcag ttgggattcg
      961 tgaattgctg ccctctggtt atgtgtggga gggctaagca cttcgtggcc gaggagcagg
     1021 actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     1081 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     1141 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     1201 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     1261 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     1321 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     1381 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     1441 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     1501 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     1561 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1621 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1681 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1741 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1801 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1861 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1921 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1981 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2041 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2101 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2161 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2221 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2281 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2341 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     2401 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     2461 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     2521 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     2581 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     2641 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     2701 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     2761 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     2821 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     2881 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     2941 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3001 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     3061 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     3121 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     3181 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     3241 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     3301 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     3361 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     3421 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     3481 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     3541 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     3601 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     3661 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
     3721 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     3781 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     3841 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     3901 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3961 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4021 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4081 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     4141 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4201 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4261 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     4321 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta
     4381 gcaccatggg acctaagaaa aagaggaagg tggcggccgc tgactacaag gatgacgacg
     4441 ataaatctag aatgggtccc gacatcgtga tgacccagag ccccagcagc ctgagcgcca
     4501 gcgtgggcga ccgcgtgacc atcacctgcc gcagcagcac cggcgccgtg accaccagca
     4561 actacgccag ctgggtgcag gagaagcccg gcaagctgtt caagggcctg atcggcggca
     4621 ccaacaaccg cgcccccggc gtgcccagcc gcttcagcgg cagcctgatc ggcgacaagg
     4681 ccaccctgac catcagcagc ctgcagcccg aggacttcgc cacctacttc tgcgccctgt
     4741 ggtacagcaa ccactgggtg ttcggccagg gcaccaaggt ggagctgaag cgcggcggcg
     4801 gcggcagcgg cggcggcggc agcggcggcg gcggcagcag cggcggcggc agcgaggtga
     4861 agctgctgga gagcggcggc ggcctggtgc agcccggcgg cagcctgaag ctgagctgcg
     4921 ccgtgagcgg cttcagcctg accgactacg gcgtgaactg ggtgcgccag gcccccggcc
     4981 gcggcctgga gtggatcggc gtgatctggg gcgacggcat caccgactac aacagcgccc
     5041 tgaaggaccg cttcatcatc agcaaggaca acggcaagaa caccgtgtac ctgcagatga
     5101 gcaaggtgcg cagcgacgac accgccctgt actactgcgt gaccggcctg ttcgactact
     5161 ggggccaggg caccctggtg accgtgagca gctacccata cgatgttcca gattacgctg
     5221 gtggaggcgg aggttctggg ggaggaggta gtggcggtgg tggttcagga ggcggcggat
     5281 ccagcggcag cgagactccc gggacctcag agtccgccac acccgaaagt atgagctcag
     5341 agactggccc agtggctgtg gaccccacat tgagacggcg gatcgagccc catgagtttg
     5401 aggtattctt cgatccgaga gagctccgca aggagacctg cctgctttac gaaattaatt
     5461 gggggggccg gcactccatt tggcgacata catcacagaa cactaacaag cacgtcgaag
     5521 tcaacttcat cgagaagttc acgacagaaa gatatttctg tccgaacaca aggtgcagca
     5581 ttacctggtt tctcagctgg agcccatgcg gcgaatgtag tagggccatc actgaattcc
     5641 tgtcaaggta tccccacgtc actctgttta tttacatcgc aaggctgtac caccacgctg
     5701 acccccgcaa tcgacaaggc ctgcgggatt tgatctcttc aggtgtgact atccaaatta
     5761 tgactgagca ggagtcagga tactgctgga gaaactttgt gaattatagc ccgagtaatg
     5821 aagcccactg gcctaggtat ccccatctgt gggtacgact gtacgttctt gaactgtact
     5881 gcatcatact gggcctgcct ccttgtctca acattctgag aaggaagcag ccacagctga
     5941 cattctttac catcgctctt cagtcttgtc attaccagcg actgccccca cacattctct
     6001 gggccaccgg gttgaaaggc ggaggtggaa gcactaatct gtcagatatt attgaaaagg
     6061 agaccggaaa gcaactggtt atccaggaat ccatcctcat gctcccagag gaggtggaag
     6121 aagtcattgg gaacaagccg gaaagcgata tactcgtgca caccgcctac gacgagagca
     6181 ccgacgagaa tgtcatgctt ctgactagcg acgcccctga atacaagcct tgggctctgg
     6241 tcatacagga tagcaacggt gagaacaaga ttaagatgct cggaggagga ggaagcggag
     6301 gaggaggtag cggaggaggt ggaagccgga ccgaagagta caagcttatc ctgaacggta
     6361 aaaccctgaa aggtgaaacc accaccgaag ctgttgacgc tgctaccgcg gaaaaagttt
     6421 tcaaacagta cgctaacgac aacggtgttg acggtgaatg gacctacgac gacgctacca
     6481 aaaccttcac ggtaaccgaa ggtggtggta gcggtggtgg tggtagtccc aagaagaaga
     6541 ggaaagtctc gagcggtgga gctgcaggag ggcccttcga aggtaagcct atccctaacc
     6601 ctctcctcgg tctcgattct acgcgtaccg gtcatcatca ccatcaccat tgagtttaaa
     6661 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6721 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6781 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6841 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     6901 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     6961 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7021 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7081 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7141 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     7201 agacggtttt tcgccctttg acgttggagt ccacg
//
