LOCUS       40924_13230        5522 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCR2.1-sacB, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5522)
  AUTHORS   Rholl D, Bruckbauer S.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-FEB-2013) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   2  (bases 1 to 5522)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 5522)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (05-FEB-2013) Microbiology Immunology and Pathology, Colorado State 
            University, IDRC at Foothills Campus, Campus Delivery 0922, Fort 
            Collins, CO 80523-0922, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5522
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(70..88)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(95..111)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      253..633
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             1001..1792
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1813..2670
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2844..3432
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3720..3741
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        3756..3786
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3794..3810
                     /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)."
     primer_bind     3818..3834
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4084..5436
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="GATQAFAKETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVSE
                     FDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQ
                     KVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGK
                     HYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHT
                     LRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKK
                     RTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGS
                     KMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKG
                     NNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK"
     terminator      complement(5453..5496)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 aagccgaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc
       61 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
      121 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      181 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      241 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
      301 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      361 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      421 ggttccgatt tagagcttta cggcacctcg accgcaaaaa acttgatttg ggtgatggtt
      481 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      541 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc gcggtctatt
      601 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      661 aacaaattca gggcgcaagg gctgctaaag gaaccggaac acgtagaaag ccagtccgca
      721 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc
      781 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc
      841 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg
      901 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg
      961 atcaagatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     1021 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     1081 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1141 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1201 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1261 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcgcct
     1321 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1381 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1441 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1501 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgat
     1561 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcaa
     1621 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg atacccgtga
     1681 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1741 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattga
     1801 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1861 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1921 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1981 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtcat
     2041 acactattat cccgtattga cgccgggcaa gagcaactcg gtcgccgggc gcggtattct
     2101 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2161 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2221 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2281 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagagt
     2341 gacaccacga tgcctgtagc aatgccaaca acgttgcgca aactattaac tggcgaacta
     2401 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2461 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2521 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2581 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2641 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2701 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2761 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2821 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2881 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2941 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     3001 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3061 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3121 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3181 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     3241 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3301 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3361 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     3421 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     3481 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     3541 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3601 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     3661 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     3721 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     3781 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     3841 tacgccaagc ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt
     3901 cggcttatgc atctagtcac ttcttgaatt tgtcaaaacc cgtccatata attagcactc
     3961 gctgcacagg aggaaacgat gacgtactcc aaacaagccg tttcccgtag acacaaacgt
     4021 ttggccctga gcgccgccgc gctcaccgca gccgcttgca tgtcggcgca cgcgcagagc
     4081 gacggcgcaa ctcaagcgtt tgcgaaagaa acgaaccaaa agccatataa ggaaacatac
     4141 ggcatttccc atattacacg ccatgatatg ctgcaaatcc ctgaacagca aaaaaatgaa
     4201 aaatatcaag tttctgaatt tgattcgtcc acaattaaaa atatctcttc tgcaaaaggc
     4261 ctggacgttt gggacagctg gccattacaa aacgctgacg gcactgtcgc aaactatcac
     4321 ggctaccaca tcgtctttgc attagccgga gatcctaaaa atgcggatga cacatcgatt
     4381 tacatgttct atcaaaaagt cggcgaaact tctattgaca gctggaaaaa cgctggccgc
     4441 gtctttaaag acagcgacaa attcgatgca aatgattcta tcctaaaaga ccaaacacaa
     4501 gaatggtcag gttcagccac atttacatct gacggaaaaa tccgtttatt ctacactgat
     4561 ttctccggta aacattacgg caaacaaaca ctgacaactg cacaagttaa cgtatcagca
     4621 tcagacagct ctttgaacat caacggtgta gaggattata aatcaatctt tgacggtgac
     4681 ggaaaaacgt atcaaaatgt acagcagttc atcgatgaag gcaactacag ctcaggcgac
     4741 aaccatacgc tgagagatcc tcactacgta gaagataaag gccacaaata cttagtattt
     4801 gaagcaaaca ctggaactga agatggctac caaggcgaag aatctttatt taacaaagca
     4861 tactatggca aaagcacatc attcttccgt caagaaagtc aaaaacttct gcaaagcgat
     4921 aaaaaacgca cggctgagtt agcaaacggc gctctcggta tgattgagct aaacgatgat
     4981 tacacactga aaaaagtgat gaaaccgctg attgcatcta acacagtaac agatgaaatt
     5041 gaacgcgcga acgtctttaa aatgaacggc aaatggtacc tgttcactga ctcccgcgga
     5101 tcaaaaatga cgattgacgg cattacgtct aacgatattt acatgcttgg ttatgtttct
     5161 aattctttaa ctggcccata caagccgctg aacaaaactg gccttgtgtt aaaaatggat
     5221 cttgatccta acgatgtaac ctttacttac tcacacttcg ctgtacctca agcgaaagga
     5281 aacaatgtcg tgattacaag ctatatgaca aacagaggat tctacgcaga caaacaatca
     5341 acgtttgcgc cgagcttcct gctgaacatc aaaggcaaga aaacatctgt tgtcaaagac
     5401 agcatccttg aacaaggaca attaacagtt aacaaataat agtagctttc aaataaaacg
     5461 aaaggctcag tcgaaagact gggcctttcg ttttaatctg agacgtccgc aattaaagat
     5521 ct
//
