LOCUS       62056_21360        6767 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 6767)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      64..158
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      650..872
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             920..1867
                     /codon_start=1
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      complement(2408..2494)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2888..4315)
                     /codon_start=1
                     /label=Tn5 transposase
                     /note="transposase from the bacterial Tn5 transposon
                     (Reznikoff, 1993)"
                     /translation="MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKS
                     ITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYR
                     HQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESG
                     KWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGL
                     YLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNA
                     VLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAER
                     QRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTP
                     DECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQS
                     KLDGFLAAKDLMAQGIKI"
     protein_bind    complement(4345..4361)
                     /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(4369..4398)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             complement(4633..5712)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5713..5790)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             join(6006..6767,1..30)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
ORIGIN
        1 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gagagctcgc
       61 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
      121 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagg gacagtaaga
      181 cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt ctgaatgacc
      241 tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa ctgctgaaca
      301 gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga gaagcccgtg
      361 acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag gcgcctgtag
      421 tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa
      481 agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg atttgcccga
      541 gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta gaggagcaaa
      601 cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg agttatacac
      661 agggctggga tctattcttt ttatcttttt ttattctttc tttattctat aaattataac
      721 cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag agggtctagc
      781 agaatttaca agttttccag caaaggtcta gcagaattta cagataccca caactcaaag
      841 gaaaaggact agtaattatc attgactagc ccatctcaat tggtatagtg attaaaatca
      901 cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg aactagcgat
      961 tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg tggcactact
     1021 caaccccacg attgaaaacc ctacaaggaa agaacggacg gtatcgttca cttataacca
     1081 atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag ctaaagcaac
     1141 cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag gctttgagat
     1201 tttccagtgg acaaactatg ccaagttctc aagcgaaaaa ttagaattag tttttagtga
     1261 agagatattg ccttatcttt tccagttaaa aaaattcata aaatataatc tggaacatgt
     1321 taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa aagaactaac
     1381 acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta agttcatgtt
     1441 aatgcttgaa aataactacc atgagtttaa aaggcttaac caatgggttt tgaaaccaat
     1501 aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc gaggccgccc
     1561 gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg taaccgaact
     1621 tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta catcagattc
     1681 ctacctacat aacggactaa gaaaaacact acacgatgct ttaactgcaa aaattcagct
     1741 caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagtatgatc tcaatggttc
     1801 gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg ctaaatacgg
     1861 aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga ctaacaaaca
     1921 aaagtagaac aactgttcac cgttacatat caaagggaaa actgtccata tgcacagatg
     1981 aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt gcattcaaag
     2041 ctgttcacca tgaacagatc gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa
     2101 caggtgaaac cagtaaaaca aagcaactag aacatgaaat tgaacacctg agacaacttg
     2161 ttacagctca acagtcacac atagacagcc tgaaacaggc gatgctgctt atcgaatcaa
     2221 agctgccgac aacacgggag ccagtgacgc ctcccgtggg gaaaaaatca tggcaattct
     2281 ggaagaaata gcgctttcag ccggcaaacc ggctgaagcc ggatctgcga ttctgataac
     2341 aaactagcaa caccagaaca gcccgtttgc gggcagcaaa acccgtaccc taggtctagg
     2401 gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     2461 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ctagagcttg catgcccaag
     2521 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     2581 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     2641 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     2701 atcgtataat ccttactgcg ttaaaggctt ttcttaggaa agttggccat ttcttaattc
     2761 agccattaat taagaaatat taagaatatt cctggctatt ttctcctgtc agagtctatt
     2821 gttttagcct gaaaagctaa aaaacgttaa caagcttgca tgcctgcagg tcgactctag
     2881 aggatccgat tttaatgccc tgcgccatca ggtctttcgc ggccagaaag ccatccagtt
     2941 tgctttgcag cgcttcccaa ccttcccaca gcgcacccca gctcgcaatg ccggtacgtt
     3001 tgctatccat aaagccgccc agacgcgcaa tcgccatata cgcccattgc aggctgcccg
     3061 ctttttcttt gcgtttgcgt ttgcctttat ccagatagcc cagcagttgg cattcatccg
     3121 gggtcagcac ggtttccgcg ctctggcttt caacgtgttc cgcttctttc agcaggccct
     3181 gcgcacgcag tgcttgcggc ggagtaaaag attcacgcag ttgcagcaga cgcaccgcca
     3241 caaagctcag aatgctcacc atacgttcca ggttatccgg ttcttccata cgctgacgtt
     3301 ccgcacccgc acccgttttc cacgctttgt gaaattcttc aatgcgccaa cgatgggtat
     3361 aaatatcaat cacacgcagc gcttgggcca gactttccac cggctcgctg gtcagcagca
     3421 gccatttcag cggggtttcg cctttcggcg gattaatttc ttcggccagc accgcgttca
     3481 gggtaatgtt gccctgtttc agggtaatac ggccgctacg caggctcagg ctcgctttac
     3541 gcgccggacg gtttttacgt ttgccacgtt tatccaccac gcctttctgc ggaatgctaa
     3601 tctgatagcc gcccagttcc ggctggtttt tcaggtgatc atacagatac aggccgcttt
     3661 ccacatcttt acgcggatgt ttgctacgca ccacaaaacg ttcgttatgg gccagtttat
     3721 cttgcagata cgcatgaata tccgcttcac gatcgcacac cgcaatcacg ttgctcatca
     3781 tgctgcccat tctcagacgc gaagttgcag cagcggccag ccatttgccg ctttcttttt
     3841 catccgcatc cgccggatca tccggacgca tccaccattc ttgatgcagc aggcccacgg
     3901 tacgaaaggt ggtcgcttcc agcagcagca cgctatgcac ccaccaacca cggcttttat
     3961 cctgaatgct acccagtttg cccagttctt ccgccacctg atgacgatag ctcagagagg
     4021 tggtatcttc aattgccagc agttccggaa attcctgggc cagtttcacg gtctgcatgg
     4081 cacccgcttt acgaatcgct tccgcgctca cgttcggatt acgaataaaa cgatacgcgc
     4141 cttcctgcat ggctttgctg ccttcgctgc taatggtaat gcttttgccg ctatatttgg
     4201 ccagttgcgc cgcaacattc accagacgcg cggtacgacg cggatcaccc agcgcagcac
     4261 tagaaaacac gcttttcgcc caatccgccg cacgatgcag tgcactggta atcatgaatt
     4321 ccatggtctg tttcctgtgt gaaattgtta tccgctcaca attccacaca ttatacgagc
     4381 cggatgatta attgtcaaca gctcatttca gaatatttgc cagaaccgtt atgatgtcgg
     4441 cgcaaaaaac attatccaga acgggagtgc gccttgagcg acacgaatta tgcagtgatt
     4501 tacgacctgc acagccatac cacagcttcc gatggctgcc tgacgccaga agcattggtg
     4561 caccgtgcag tcgatgataa gctgtcaaac cagatcaatt cgcgctaact cacattaatt
     4621 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4681 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     4741 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     4801 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttgacgg
     4861 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     4921 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     4981 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     5041 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     5101 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     5161 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     5221 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     5281 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     5341 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     5401 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     5461 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     5521 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     5581 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     5641 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     5701 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     5761 gcgaaaggtt ttgcaccatt cgatggtgtc cgaggtgaag acgaaagggc ctcgtgatac
     5821 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtcg gaattgccag
     5881 ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt aaactggatg gctttcttgc
     5941 cgccaaggat ctgatggcgc aggggatcaa gatctgatca agagacagga tgaggatcgt
     6001 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     6061 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     6121 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     6181 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     6241 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     6301 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     6361 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     6421 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     6481 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     6541 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     6601 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     6661 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     6721 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatc
//
