LOCUS       40924_35693        4596 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPV570, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4596)
  AUTHORS   Lok JB, Shao H, Massey HC, Li X.
  TITLE     Transgenesis in Strongyloides and related parasitic nematodes: 
            historical perspectives, current functional genomic applications and
            progress towards gene disruption and editing
  JOURNAL   Parasitology (2016) In press
  PUBMED    27000743
REFERENCE   2  (bases 1 to 4596)
  AUTHORS   Lok JB, Shao H, Massey HC Jr., Li X.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2016) Pathobiology, University of Pennsylvania, 
            3800 Spruce Street, Philadelphia, PA 19104, USA
REFERENCE   3  (bases 1 to 4596)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4596)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Parasitology (2016) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2016) Pathobiology, University of Pennsylvania, 3800 Spruce 
            Street, Philadelphia, PA 19104, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..128
                     /label=AmpR promoter
     CDS             129..986
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1160..1748
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2956..3467
                     /note="U6; Strogyloides ratti promoter"
                     /regulatory_class="promoter"
     misc_feature    3467..3489
                     /label=gRNA for Cherry target
                     /note="gRNA for Cherry target"
     misc_RNA        3492..3567
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     3576..3592
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3981..3997)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      4139..4594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
       61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
      541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
      601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
      661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
      721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
      781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
      841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
      901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
      961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     1801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     1861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     1921 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     1981 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     2041 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     2101 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     2161 ccaagcgcgc ccgccgggta actcacgggg tatccatgtc catttctgcg gcatccagcc
     2221 aggatacccg tcctcgctga cgtaatatcc cagcgccgca ccgctgtcat taatctgcac
     2281 accggcacgg cagttccggc tgtcgccggt attgttcggg ttgctgatgc gcttcgggct
     2341 gaccatccgg aactgtgtcc ggaaaagccg cgacgaactg gtatcccagg tggcctgaac
     2401 gaacagttca ccgttaaagg cgtgcatggc cacaccttcc cgaatcatca tggtaaacgt
     2461 gcgttttcgc tcaacgtcaa tgcagcagca gtcatcctcg gcaaactctt tccatgccgc
     2521 ttcaacctcg cgggaaaagg cacgggcttc ttcctccccg atgcctggat agcgccagct
     2581 tgggcgatga ctgagccgga aaaaagaccc gacgatatga tcctgatgca gctagattaa
     2641 ccctagaaag atagtctgcg taaaattgac gcatgcattc ttgaaatatt gctctctctt
     2701 tctaaatagc gcgaatccgt cgctgtgcat ttaggacatc tcagtcgccg cttggagctc
     2761 ccgtgaggcg tgcttgtcaa tgcggtaagt gtcactgatt ttgaactata acgaccgcgt
     2821 gagtcaaaat gacgcatgat tatcttttac gtgactttta agatttaact catacgataa
     2881 ttatattgtt atttcatgtt ctacttacgt gataacttat tatatatata ttttcttgtt
     2941 atagatatca agcttgttcc aatagaacat aaaggaaaaa tttatcaagc tacatatact
     3001 gatctttgta tgacatatga ttggaaatgt tttcttaatg atcatgttga tgttttatta
     3061 ccaaaagaac aacatttagg tacatttaaa ggtatgtata atattaaaat atttttattt
     3121 attacatgta attaatagta gaaatatttt ttataacttt ttattttact tttactaggg
     3181 atatccctta ttacattaga taaaatattt atacatacat ttacagtgta aatttttatt
     3241 aaatattata atatttaaat tatttttaaa aaaaatatta aaaaatataa tttaatttta
     3301 atagagataa tttttttaat atattatata tataaaaata taactttatt ttacattaca
     3361 aactaacata tacatacata atttcttatt caacccggct aatatagtgg agagtaaaaa
     3421 cggagtgtta tataaaaaag attgccatcc aatacaatgt tanccggtta aaggagagga
     3481 agacaacatg ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
     3541 cttgaaaaag tggcaccgag tcggtgcttt ttttntctag aactagtgga tcccccgggc
     3601 tgcaggaatt cgataaaagt tttgttactt tatagaagaa attttgagtt tttgtttttt
     3661 tttaataaat aaataaacat aaataaattg tttgttgaat ttattattag tatgtaagtg
     3721 taaatataat aaaacttaat atctattcaa attaataaat aaacctcgat atacagaccg
     3781 ataaaacaca tgcgtcaatt ttacgcatga ttatctttaa cgtacgtcac aatatgatta
     3841 tctttctagg gttaatctag ctgcgtgttc tgcagcgtgt cgagcatctt catctgctcc
     3901 atcacgctgt aaaacacatt tgcaccgcga gtctgcccgt cctccacggg ttcaaaaacg
     3961 tgaatgaacg aggcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     4021 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4081 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
     4141 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     4201 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     4261 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     4321 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     4381 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     4441 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     4501 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     4561 gcgaatttta acaaaatatt aacgcttaca atttag
//
