LOCUS       62056_19195        6973 bp DNA     circular SYN 02-APR-2019
DEFINITION  Cloning vector pSAS31_redac, complete sequence.
ACCESSION   MK178599
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6973)
  AUTHORS   Scholz SA, Diao R, Wolfe MB, Fivenson EM, Lin XN, Freddolino PL.
  TITLE     High-Resolution Mapping of the Escherichia coli Chromosome Reveals 
            Positions of High and Low Transcription
  JOURNAL   Cell Syst (2019) In press
  PUBMED    30904377
REFERENCE   2  (bases 1 to 6973)
  AUTHORS   Scholz SA.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-NOV-2018) Biological Chemistry, University of 
            Michigan, 1150 W. Medical Center Dr., Ann Arbor, MI 48109, USA
REFERENCE   3  (bases 1 to 6973)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Syst 
            (2019) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (15-NOV-2018) Biological Chemistry, University of Michigan, 1150 W. 
            Medical Center Dr., Ann Arbor, MI 48109, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6973
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(791..933)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(1038..1226)
                     /codon_start=1
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
                     /translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     protein_bind    complement(2001..2022)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(2038..3117)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(3118..3195)
                     /label=lacI promoter
     misc_feature    3711..3729
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     CDS             complement(3855..4646)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     protein_bind    complement(5014..5047)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     protein_bind    5060..5078
                     /label=tet operator
                     /note="bacterial operator O2 for the tetR and tetA genes"
     gap             5141..5160
                     /estimated_length=20
     misc_feature    complement(5274..5292)
                     /label=Tn5 ME
                     /note="hyperactive mosaic end for Tn5 transposase
                     recognition (Reznikoff et al., 2004)"
     terminator      5421..5468
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5505..5960
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(6056..6868)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      661 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      721 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      781 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      841 tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
      901 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      961 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     1021 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     1081 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     1141 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     1201 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     1261 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     1321 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     1381 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     1441 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     1501 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     1561 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     1621 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     1681 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     1741 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     1801 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     1861 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     1921 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     1981 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     2041 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     2101 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     2161 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     2221 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     2281 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     2341 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2401 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2461 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2521 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2581 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2641 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2701 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2761 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2821 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2881 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2941 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     3001 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     3061 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     3121 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     3181 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     3241 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     3301 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     3361 cgctcatctc tcccttatgc gactcctgca ttaggaagca gcccagtagt aggttgaggc
     3421 cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc aacagtcccc
     3481 cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc
     3541 gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg
     3601 cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg aagcttcatg
     3661 ctgtaaaaag catgataata aattaaaagc gatgtaaata atttatgcag ctgtctctta
     3721 tacacatctc ctcggtacca aattccagaa aagaggcctc ccgaaagggg ggcctttttt
     3781 cgttttggtc cgaagttcct atactttcta gagaatagga acttcggaat aggaacttca
     3841 agatcccctt attagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     3901 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     3961 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     4021 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     4081 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     4141 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     4201 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     4261 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     4321 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     4381 cttcccgctt cagtgacaac gtcgagcact gctgcgcaag gaacgcccgt cgtggccagc
     4441 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     4501 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     4561 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     4621 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     4681 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag
     4741 ggcttcccaa ccttaccaga gggcgcccca gcaggcaatt ccggttcgct tgctgtccat
     4801 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt
     4861 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     4921 cgtttctgcg gactggcttt ctacgtgttc cgcttccttt agcagccctt gcgccctgag
     4981 tgcttgcggc agcgtgagct tcaaaagcgc tctgaagttc ctatactttc tagagaatag
     5041 gaacttcttt atagctagct ccctatcagt gatagagatt gacatcccta tcagtgatag
     5101 agatactgag cacacataga ttaaagagga gaaattacat nnnnnnnnnn nnnnnnnnnn
     5161 taatacgact cactatagac actctttccc tacacgacgc tcttccgatc tggcgcgcct
     5221 accaccgtca aaaaaaacgg cgctttttag cgccgttttt atttttcaac cttagatgtg
     5281 tataagagac agctgcacaa agttatccac atgacgattt gcgagcgatc cagaagatct
     5341 acaaaagaat tcactgagat ccggctgcta acaaagcccg aaaggaagct gagttggctg
     5401 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     5461 gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc cctgtagcgg
     5521 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5581 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5641 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5701 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5761 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5821 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5881 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     5941 aatattaacg tttacaattt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6001 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     6061 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     6121 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     6181 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     6241 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     6301 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     6361 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     6421 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     6481 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     6541 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     6601 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     6661 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     6721 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     6781 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     6841 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     6901 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6961 gaccccgtag aaa
//
