LOCUS       40924_35678        8024 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pPV512, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8024)
  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 8024)
  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 8024)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8024)
  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..8024
                     /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"
     CDS             complement(3315..3326)
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     misc_feature    4259..5124
                     /label=similar to green fluorescent protein
                     /note="similar to green fluorescent protein"
     CDS             5132..5602
                     /label=ecDHFR
                     /note="E. coli dihydrofolate reductase"
     CDS             5603..5629
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    5629..5716
                     /note="P2A; linker"
     CDS             5648..5704
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             5717..6391
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     misc_feature    6428..6990
                     /note="Era1; UTR"
     primer_bind     7004..7020
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7409..7425)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      7567..8022
                     /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 agcttgatta tcgtagtgag aggacgtgta gagataataa atttcccaac
     3001 ttaatctatt ataggaactt agttaaaaat gattgtattg gtcctctttg tgaaagtgtg
     3061 gaaaagaggt acctcgtagt ttttttcttc tcaattctca tcatcaccct gtgtaacatc
     3121 acatctcata tagagaaatc gcgactttta ttgcttagca tatactttgt aagttgtttt
     3181 cactgaggct ttgtccatca cacgctacac attatcgctg ttatttttat tatttatatt
     3241 tattagtaat aatattaaat aaacacatag gaattttgtc tttgttggcc cttccagcct
     3301 aaatagttat aaaatctacc ttctattata catacgcata tgttatttaa aaagttgaag
     3361 tgatttctct tctacagaaa ctacacattt atcttcttat tttcttccaa acatatagca
     3421 ataataattt aaaacttttt cttactcctc ccagagtttg ttaagttgat catatttaaa
     3481 attcttctgt cttctacaac aaataatata tttattataa caccaactac ctcgttccaa
     3541 gtattacgta ggagcatata cataccacaa tgcttaggaa atgggggcgg ttatgatata
     3601 acattatttc aatgtaacat tcttttttac cacatatatt ttctactgaa ttctgaaaac
     3661 tcaatttcaa acctcctata tatatatatt gttaggcact tctaagatta taattcattt
     3721 aactctctta aatttatcta ttttaaattt ttcctttttt gtattattta tactattata
     3781 tcgaaacttt attattacat gtactattta acttttattt tcttatataa taataatact
     3841 aaataataac tgataatcgt atctgtagag attattgaat tgtgattagt aatgtaaagt
     3901 aaatctatat taagatatct caaagatgca ctcaagttat ttgaaaaatt tcttatacat
     3961 agattaatac ttatattaag aaactttaaa aaaaagtttt aaataaaaaa aaaaattcac
     4021 aattgtacta ttttatcttg tcgttcagtt tatcaccgta gtatttttat ggctagtaat
     4081 attcatcatt tttagataaa ctaacttgct atagaaaatg tgtgacgaag aggttgccgc
     4141 tgcaggtcga ctagaggatc cccgggattg gccaaaggac ccaaaggtat gtttcgaatg
     4201 atactaacat aacatagaac attttcagga ggacccttgg agggtaccgg tagaaaaaat
     4261 gagtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga
     4321 tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa
     4381 acttaccctt aaatttattt gcactactgg aaaactacct gttccatggg taagtttaaa
     4441 catatatata ctaactaacc ctgattattt aaattttcag ccaacacttg tcactacttt
     4501 ctgttatggt gttcaatgct tctcgagata cccagatcat atgaaacggc atgacttttt
     4561 caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca aagatgacgg
     4621 gaactacaag acacgtaagt ttaaacagtt cggtactaac taaccataca tatttaaatt
     4681 ttcaggtgct gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg
     4741 tattgatttt aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc
     4801 acacaatgta tacatcatgg cagacaaaca aaagaatgga atcaaagttg taagtttaaa
     4861 catgatttta ctaactaact aatctgattt aaattttcag aacttcaaaa ttagacacaa
     4921 cattgaagat ggaagcgttc aactagcaga ccattatcaa caaaatactc caattggcga
     4981 tggccctgtc cttttaccag acaaccatta cctgtccaca caatctgccc tttcgaaaga
     5041 tcccaacgaa aagagagacc acatggtcct tcttgagttt gtaacagctg ctgggattac
     5101 acatggcatg gatgaactat acaatccgga cagtctgatt gcggcgttag cggtagatca
     5161 cgttatcggc atggaaaccg tcatgccgtg gaacctgcct gccgatctcg cctggtttaa
     5221 acgcaacacc ttaaataaac ccgtgattat gggccgccat acctgggaat caatcggtcg
     5281 tccgttgcca ggacgcaaaa atattatcct cagcagtcaa ccgagtacgg acgatcgcgt
     5341 aacgtgggtg aagtcggtgg atgaagccat cgcggcgtgt ggtgacgtac cagaaatcat
     5401 ggttattggc ggcggtcgcg tttatgaaca gttcttgcca aaagcgcaaa aactgtatct
     5461 gacgcatatc gacgcagaag tggaaggcga cacccatttc ccggattacg agccggatga
     5521 ctgggaatcg gtattcagcg aattccacga tgctgatgcg cagaactctc acagctattg
     5581 ctttgagatt ctggagcggc gatacccata cgatgttcca gattacgctt ggctagccgg
     5641 aagcggagct actaacttca gcctgctgaa gcaggctgga gacgtggagg agaaccctgg
     5701 acctcggccg gtaaaaatgg catcatcaga agatgttatt aaagaattta tgagattcaa
     5761 agttaaaatg gaaggttcag ttaacggtca tgaatttgaa attgaaggtg aaggtgaagg
     5821 tagaccatat gaaggtacac aaacagcaaa attaaaagtt acaaaaggtg gtccattacc
     5881 atttgcatgg gatattcttt caccacaatt tcaatatggt agtaaagcat atgttaaaca
     5941 tccagcagat attccagatt atcttaaact ttcatttcca gaaggtttta aatgggaaag
     6001 aattatgaat tttgaagatg gtggtgttgt aacagttact caagattcaa cattacaaga
     6061 tggtgaattt atttataaag ttaaattaag aggtaccaat tttccatcag atggtccagt
     6121 aatgcaaaaa aaaactatgg gttgggaagc atcaacagaa agaatgtatc cagaagatgg
     6181 tgcactaaaa ggtgaaatta aaatgagatt aaaacttaag gatggtggtc attatgatgc
     6241 agaagttaaa acaacttata aagctaaaaa acaagttcaa ttaccaggtg cttataaaac
     6301 agatattaaa ttagatatta tttcacataa tgaagattat actattgttg aacaatatga
     6361 aagagcagaa ggtagacatt caacaggtgc aggatctcct aggaatttat tttgataagt
     6421 ctcgcgtatt ttatttgtaa taatcttttt ataagatcaa tttacattta tactgataaa
     6481 attgacacaa ttacactttt tattataaaa tattttaaaa tatttctatc aaaaggttta
     6541 tttttgtttt aaatataatg gttttttttg taaaaaatac ttatttcgtt tgataatctt
     6601 tattcagaga tttgaataat tttaaataaa gtagtttagt tttcatataa aagttaatgg
     6661 tatatataaa tatttaaata gtaaagattg aatcagtatt caaaatgaca cctaaaagtt
     6721 attctatttc tagctataac aataataata aagataatta tgatacagat ttagaaaaaa
     6781 aaataaggtt aaattaatta attctatttt taattaaaat gatttaaggg agttaaatga
     6841 aacagaaaca gattgggtct ctatatattt agtttatgta atgttatttt ttggtggaat
     6901 acaaataacg gtatatttta cttcaatgtg gccatactta tcaacaataa atagtaaggc
     6961 aaatgtagat actttagggt gggttgtagc actagtcggc cgctctagaa ctagtggatc
     7021 ccccgggctg caggaattcg ataaaagttt tgttacttta tagaagaaat tttgagtttt
     7081 tgtttttttt taataaataa ataaacataa ataaattgtt tgttgaattt attattagta
     7141 tgtaagtgta aatataataa aacttaatat ctattcaaat taataaataa acctcgatat
     7201 acagaccgat aaaacacatg cgtcaatttt acgcatgatt atctttaacg tacgtcacaa
     7261 tatgattatc tttctagggt taatctagct gcgtgttctg cagcgtgtcg agcatcttca
     7321 tctgctccat cacgctgtaa aacacatttg caccgcgagt ctgcccgtcc tccacgggtt
     7381 caaaaacgtg aatgaacgag gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg
     7441 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     7501 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     7561 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     7621 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     7681 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     7741 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     7801 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     7861 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     7921 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     7981 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttag
//
