LOCUS       62056_20935        6742 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 6742)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6742
                     /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"
     promoter        complement(4345..4373)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             complement(4608..5687)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(5688..5765)
                     /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(5981..6742,1..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 gaaacattat acgagccgat gattaattgt caacagctca
     4381 tttcagaata tttgccagaa ccgttatgat gtcggcgcaa aaaacattat ccagaacggg
     4441 agtgcgcctt gagcgacacg aattatgcag tgatttacga cctgcacagc cataccacag
     4501 cttccgatgg ctgcctgacg ccagaagcat tggtgcaccg tgcagtcgat gataagctgt
     4561 caaaccagat caattcgcgc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4621 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4681 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
     4741 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
     4801 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
     4861 gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg
     4921 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
     4981 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
     5041 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
     5101 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
     5161 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
     5221 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
     5281 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
     5341 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     5401 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     5461 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     5521 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     5581 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     5641 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     5701 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgca ccattcgatg
     5761 gtgtccgagg tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5821 gataataatg gtttcttaga cgtcggaatt gccagctggg gcgccctctg gtaaggttgg
     5881 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
     5941 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     6001 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     6061 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     6121 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     6181 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     6241 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     6301 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     6361 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     6421 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     6481 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     6541 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     6601 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     6661 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     6721 cgctcccgat tcgcagcgca tc
//
