LOCUS       Exported                4648 bp ds-DNA     circular SYN 04-MAR-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4648)
  AUTHORS   Blodgett lab WUSTL
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jan 8, 2020 from SnapGene 5.0.7
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4648
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(2..43)
                     /label=pJMD14L_pJMD17_rev
                     /note="pJMD9_pJMD14_fwd
                     JMD115"
                     /note="color: black; sequence: 
                     AGCACCTGAAGTtttggtcatgagatgactgatgtcatcagc"
     primer_bind     complement(2..43)
                     /label=pJMD9_pJMD14_rev
                     /note="pJMD14L_pJMD17_rev
                     JMD115"
                     /note="color: black; sequence: 
                     AGCACCTGAAGTtttggtcatgagatgactgatgtcatcagc"
     primer_bind     19..55
                     /label=pJMD14S_pJMD17_fwd
                     /note="pRARE2_pJMD14_revJMD114"
                     /note="color: black; sequence: 
                     tctcatgaccaaaACTTCAGGTGCTACATTTGAAGAG"
     primer_bind     19..55
                     /label=pRARE2_pJMD14_rev
                     /note="JMD114"
                     /note="color: black; sequence: 
                     tctcatgaccaaaACTTCAGGTGCTACATTTGAAGAG"
     rep_origin      109..653
                     /direction=RIGHT
                     /label=p15A oriR
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
                     /note="color: #ffff00"
     primer_bind     complement(204..225)
                     /label=JMD110
                     /note="p15A_seq_"
                     /note="color: black; sequence: AGCCAGTTACCTCGGTTCAAAG"
     primer_bind     460..481
                     /label=JMD109
                     /note="p15Aseq_fwd"
                     /note="color: black; sequence: TCAGGCGTGGAATGAGACAAAC"
     primer_bind     593..612
                     /label=BK MCS/terminator seq for
                     /note="color: black; sequence: CCACCACTGATTTGAGCGTC"
     primer_bind     complement(690..722)
                     /label=pRARE2_pJMD14_fwd
                     /note="JMD113"
                     /note="color: black; sequence: 
                     aatgaatcggccTGGAAGATGCCAGGAAGATAC"
     primer_bind     698..731
                     /label=pJMD9_pJMD14_fwd
                     /note="JMD116"
                     /note="color: black; sequence: 
                     CTGGCATCTTCCAggccgattcattaatgcagct"
     primer_bind     711..730
                     /label=PXS6
                     /note="color: black; sequence: GGCCGATTCATTAATGCAGC"
     misc_feature    798..1155
                     /label=pErmE* cloning cassette
                     /note="color: #a6acb3"
     terminator      798..828
                     /label=fd terminator
                     /label=fd-ter
                     /note="color: #ffffff; direction: BOTH"
     misc_feature    829..901
                     /label=MCS
                     /note="pBluescript multiple cloning site"
                     /note="color: #99ccff"
     promoter        complement(join(912..917,958..963,978..983,1026..1031,
                     1050..1055))
                     /label=pErmE* promoter (pDA1652)
                     /note="This reverse directional feature has 5 segments:
                        1:  912 ..  917 / #999999 / RBS
                        2:  958 ..  963 / #ffffff / -10 1
                        3:  978 ..  983 / #ffffff / -35 1
                        4: 1026 .. 1031 / #ffffff / -10 2
                        5: 1050 .. 1055 / #ffffff / -35 2"
     terminator      1061..1155
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff"
     primer_bind     1200..1219
                     /label=YQ325-pJMD9-F
                     /note="color: black; sequence: CACGCCCTCCTACATCGAAG"
     primer_bind     1200..1219
                     /label=phiOZJ_CK_att_P
                     /note="YQ325-pJMD9-F"
                     /note="color: black; sequence: CACGCCCTCCTACATCGAAG"
     primer_bind     complement(1202..1220)
                     /label=pJMD14S_pJMD17_rev
                     /note="JMD167"
                     /note="color: black; sequence: 
                     ATGCTCCCGCTCTCGcttcgatgtaggaggGCG"
     primer_bind     complement(1206..1235)
                     /label=pJMD14_RDF_del_fwd
                     /note="color: black; sequence: 
                     GATATCGACGTCGCTTTCAGCTTCGATGTAGGAG"
     primer_bind     1227..1251
                     /label=pJMD14_RDF_del_rev
                     /note="color: black; sequence: 
                     GATATCGACGTCGAGAGAGGCGAGGAGAGC"
     misc_feature    1234..1317
                     /label=Partial Phi-OZJ RDF
                     /note="color: #993300"
     misc_feature    complement(1314..1410)
                     /label=B12 binding protein C-terminus
                     /note="color: #000080; direction: LEFT"
     primer_bind     complement(1336..1355)
                     /label=BK MCS/term seq rev
                     /note="color: black; sequence: CTCTCGGTCAAGAAGCCCTC"
     primer_bind     1350..1368
                     /label=pJMD14L_pJMD17_fwd
                     /note="JMD166"
                     /note="color: black; sequence: 
                     cctcctacatcgaagCGAGAGCGGGAGCATCTTC"
     misc_feature    1384..1433
                     /label=Phi-OZJ attP
                     /note="This forward directional feature has 3 segments:
                        1: 1384 .. 1407 / #008080 / P
                        2: 1408 .. 1409 / #ffffff / O
                        3: 1410 .. 1433 / #008080 / P'"
     CDS             1452..3029
                     /codon_start=1
                     /label=Phi-OZJ int
                     /note="color: #ff6600"
                     /translation="MSARDYDIEAEWTPADLALLKELEEAEALLPADAPRALLSVRLSV
                     FTDDTTSPVRQELDLRQLAREKGHRVVGLASDLNVSATKVPPWKRKSLGDWLNNRAPEF
                     DALLFWKIDRFIRNLNDLNVMIRWSETYSKNLISKNDPIDLTTTMGKMMVSLLGGVAEI
                     EAANTKTRVESLWDYTKTQGEWHVGKPPFGYKTGRDAAGKVVLVEDPPAVETLHTAREL
                     VMSGMSTTAAAKELKERGLISSTTATLTRRLRNPGILGLRVEEDKDGGIRRSKLILGRD
                     GQPIRIADPIFTEEQFEELQAVLDKRGKRQPHRQPGGATSFLGVLKCAECGTNMINHFT
                     RNRHGDYAYLRCQGCKSGGYGAPNPQEVYDRLVEQVLAVLGDFPVEMREYARGEEKRKE
                     LKRLEESIAYYMKELEPGGRFTKTRFTQDQAEGTLDKLIAELEAIDPEAAKDRWVYVAG
                     GKTFREHWEEGGIDAMSADLIRAGIQCQVTRTKVPKVRAPQVHLKLMIPKDVRTRLVIR
                     PDDFGQTF"
     primer_bind     complement(1728..1747)
                     /label=YQ324-pJMD9-intR
                     /note="color: black; sequence: TTGTTCAACCAGTCGCCGAG"
     primer_bind     complement(1728..1747)
                     /label=phiOZJ_CK_att_P'
                     /note="YQ324-pJMD9-intR"
                     /note="color: black; sequence: TTGTTCAACCAGTCGCCGAG"
     primer_bind     1778..1799
                     /label=phiOZJ_CK_int_fwd
                     /note="JMD108"
                     /note="color: black; sequence: CTGGAAGATCGACCGGTTCATC"
     primer_bind     complement(2321..2342)
                     /label=phiOZJ_CK_int_rev
                     /note="JMD107"
                     /note="color: black; sequence: GAACTGCTCCTCGGTGAAGATC"
     primer_bind     complement(3017..3051)
                     /label=pAdB02_pJMD9_rev
                     /note="JMD098"
                     /note="color: black; sequence: 
                     ctacacgaaccctAGAGGATCCTCAGAAGGTCTGT"
     primer_bind     3026..3061
                     /label=pBTC034_pJMD9_rev
                     /note="JMD097"
                     /note="color: black; sequence: 
                     CTGAGGATCCTCTagggttcgtgtagactttccttg"
     oriT            3083..3192
                     /label=incP oriT
                     /note="incP origin of transfer"
                     /note="color: #ffff00"
     regulatory      3491..3540
                     /regulatory_class="promoter"
                     /note="aminoglycoside phosphotransferase promoter region"
                     /note="color: #ffffff; direction: RIGHT"
                     /experiment="Beck E, Ludwig G, Auerswald EA, Reiss B, 
                     Schaller H.
                     Nucleotide sequence and exact localization of the neomycin 
                     phosphotransferase gene from transposon Tn5
                     Gene 19 (3), 327-336 (1982)
                     PubMed ID:  6295884
                     "
     CDS             3629..4423
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #00ff00"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     3680..3699
                     /label=kan_CHECK_rv
                     /note="color: black; sequence: aggctattcggctatgactg"
     primer_bind     3680..3699
                     /label=kan_ck_rv
                     /note="color: black; sequence: aggctattcggctatgactg"
     primer_bind     complement(3683..3704)
                     /label=JMD094
                     /note="color: black; sequence: gtgcccagtcatagccgaatag"
     primer_bind     complement(4241..4260)
                     /label=kan_CHECK_fw
                     /note="color: black; sequence: cggccacagtcgatgaatcc"
     primer_bind     complement(4241..4260)
                     /label=kan_ck_fw
                     /note="color: black; sequence: cggccacagtcgatgaatcc"
     primer_bind     4272..4292
                     /label=JMD093
                     /note="color: black; sequence: cggaccgctatcaggacatag"
     primer_bind     complement(join(4645..4648,1..32))
                     /label=pBTC034_pJMD9_fwd
                     /note="JMD096"
                     /note="color: black; sequence: 
                     ttttggtcatgagatgactgatgtcatcagcggtgg"
ORIGIN
        1 cgctgatgac atcagtcatc tcatgaccaa aacttcaggt gctacatttg aagagataaa
       61 ttgcactgaa atctagaaat attttatctg attaataaga tgatcttctt gagatcgttt
      121 tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc
      181 gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc
      241 accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc
      301 taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact
      361 caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac
      421 agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa
      481 cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg
      541 agggagccgc caggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccaccact
      601 gatttgagcg tcagatttcg tgatgcttgt caggggggcg gagcctatgg aaaaacggct
      661 ttgccgcggc cctctcactt ccctgttaag tatcttcctg gcatcttcca ggccgattca
      721 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gccggtcaag
      781 taaggtctgg aggaacaatt aaaggctcct tttggagcct ttttttttcg aaggtaccct
      841 cgagcagatc ttgtacagtt aacgaattcg atatcactag ttctagagcg gccgcgagct
      901 ccatatgtcc gcctcctttg gtcactcagt cagatctggt actgcttaat taaccggatc
      961 ctaccaaccg gcacgattgt gcccacaaca gcatcgcggt gccacgtgtg gaccgcgtcg
     1021 gtcagatcct ccccgcacct ctcgccagcc gtcaagacct attctcacca ataaaaaacg
     1081 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
     1141 ctatcaacag gagtcagact cctcctctca acgactatag tttacccgca ggacatatcc
     1201 acgccctcct acatcgaagc tgaaagcgac gtcgagagag gcgaggagag cttccctctc
     1261 cccctcgggc agggccatca ggctctccgc caggcgctcg accacgtcgc gcttcatgcg
     1321 cgttccagca ggtcagaggg cttcttgacc gagagcggga gcatcttccc gccaggcggt
     1381 gccagttgtg ggcttgttcc catctggggg cagatggaac cacgcctaca tccagtagta
     1441 ccctgctcac catgagtgcg cgcgactacg acatcgaagc tgagtggacc ccggccgacc
     1501 tcgccctgct gaaggagctg gaggaagccg aggctctgct accagccgac gcgcctcgcg
     1561 cgctcctgtc cgtgcgcctg tccgtcttca ctgacgacac cacgtcccca gtgcggcagg
     1621 agctggatct ccgccagctt gcccgagaga agggccaccg agtcgtcggc ttggccagcg
     1681 acctgaacgt gtcggctacc aaggtaccgc catggaagcg caagtccctc ggcgactggt
     1741 tgaacaaccg agcgcccgag ttcgacgcgc tcctgttctg gaagatcgac cggttcatcc
     1801 gcaacctcaa cgacctgaac gtgatgatcc gttggtcgga gacctactcg aagaacctga
     1861 tctcgaagaa cgacccgatc gacctgacga cgaccatggg caagatgatg gtctcgctcc
     1921 tcggtggcgt cgcagagatc gaggcagcca acacgaagac ccgcgtcgag tccctgtggg
     1981 actacaccaa gacgcagggc gagtggcacg tcggcaagcc tccgttcggc tacaagacgg
     2041 ggcgtgacgc ggcgggcaag gtggtcctgg tcgaggaccc tcctgcggtc gagacgctgc
     2101 acacggcgcg cgagctggtg atgagcggga tgtccacgac ggccgccgcg aaggagctga
     2161 aggagcgcgg actgatctcg tccacgactg ccaccctgac ccgtcgcctg cgcaacccag
     2221 gcatcctggg tctccgtgtc gaggaggaca aggacggtgg catccgtcgc tcgaagctga
     2281 tcctggggcg cgacggccaa ccgatccgca tcgccgatcc gatcttcacc gaggagcagt
     2341 tcgaggagct gcaagccgtc ctcgacaagc gagggaagcg ccagcctcat cgccagccgg
     2401 gcggagcgac gagcttcctc ggggtgctga agtgcgcgga gtgcgggacg aatatgatca
     2461 accacttcac gcgcaaccgg cacggtgact acgcctacct tcggtgccag ggctgcaagt
     2521 ccggagggta cggcgcaccc aacccccagg aggtgtacga ccggctggtc gagcaggtgc
     2581 tcgctgtgct cggagacttc cccgtcgaga tgcgggagta cgcccgaggc gaggagaagc
     2641 gcaaggagct gaagcgcctg gaggagtcga tcgcctacta catgaaggag ctggagccgg
     2701 gcggtcggtt caccaagacg cggttcaccc aggaccaggc cgagggcacg ctcgacaagc
     2761 tgatcgccga gctggaagcc atcgaccccg aggcggcgaa ggaccggtgg gtctacgtcg
     2821 ccgggggcaa gacgttccgc gagcactggg aggagggcgg catcgacgcc atgtcggccg
     2881 acctgatccg ggcgggtatc cagtgccagg tgacgcggac gaaggtcccg aaggtgcgcg
     2941 caccgcaggt gcacctgaag ctaatgatcc cgaaggacgt tcggacccgc ctggtcattc
     3001 ggccagacga cttcggacag accttctgag gatcctctag ggttcgtgta gactttcctt
     3061 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
     3121 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
     3181 gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa gcggatggct gatgaaacca
     3241 agccaaccag gaagggcagc ccacctatca aggagcttca cgctgccgca agcactcagg
     3301 gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga aacggtgctg
     3361 accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag
     3421 aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac
     3481 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
     3541 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
     3601 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3661 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3721 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3781 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3841 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3901 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3961 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4021 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4081 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4141 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4201 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4261 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4321 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4381 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4441 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4501 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4561 cgcggggatc tcatgctgga gttcttcgcc caccccgggt accgagctcg gcttttcgcc
     4621 attcgtattg cacgacattg cactccac
//
