LOCUS       40924_25516        7506 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pIndigoBAC-5, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7506)
  AUTHORS   EPICENTRE Biotechnologies.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-AUG-2007) 726 Post Road, Madison, WI 53713, USA
REFERENCE   2  (bases 1 to 7506)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7506)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (23-AUG-2007) 726 Post Road, Madison, WI 53713, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     288..304
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        311..329
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    332..388
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(431..447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(455..471)
                     /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)."
     promoter        complement(479..509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(524..545)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(768..1424)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(1425..1527)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      2454..2673
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             2764..3516
                     /codon_start=1
                     /label=repE
                     /note="replication initiation protein for the bacterial F 
                     plasmid"
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    3522..3772
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             4098..5270
                     /codon_start=1
                     /label=sopA
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             5273..6241
                     /codon_start=1
                     /label=sopB
                     /note="partitioning protein for the bacterial F plasmid"
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    6317..6790
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     misc_feature    complement(7050..7448)
                     /label=cos
                     /note="lambda cos site; allows packaging into phage lambda 
                     particles"
     protein_bind    complement(7466..7499)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gcggccgcaa ggggttcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
       61 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
      121 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt cagctgcgca actgttggga
      181 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
      241 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
      301 cagtgaattg taatacgact cactataggg cgaattcgag ctcggtaccc ggggatcctc
      361 tagagtcgac ctgcaggcat gcaagcttga gtattctata gtctcaccta aatagcttgg
      421 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
      481 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
      541 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
      601 attaatgaat cggccaacgc gaaccccttg cggccgcccg ggccgtcgac caattctcat
      661 gtttgacagc ttatcatcga atttctgcca ttcatccgct tattatcact tattcaggcg
      721 tagcaaccag gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc
      781 cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa
      841 acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat
      901 ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa
      961 ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta
     1021 gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac
     1081 tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg
     1141 aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc
     1201 atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa
     1261 aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc
     1321 tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat
     1381 tgggatatat caacggtggt atatccagtg atttttttct ccattttagc ttccttagct
     1441 cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga
     1501 aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc
     1561 ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca
     1621 ggtatttatt cgcgataagc tcatggagcg gcgtaaccgt cgcacaggaa ggacagagaa
     1681 agcgcggatc tgggaagtga cggacagaac ggtcaggacc tggattgggg aggcggttgc
     1741 cgccgctgct gctgacggtg tgacgttctc tgttccggtc acaccacata cgttccgcca
     1801 ttcctatgcg atgcacatgc tgtatgccgg tataccgctg aaagttctgc aaagcctgat
     1861 gggacataag tccatcagtt caacggaagt ctacacgaag gtttttgcgc tggatgtggc
     1921 tgcccggcac cgggtgcagt ttgcgatgcc ggagtctgat gcggttgcga tgctgaaaca
     1981 attatcctga gaataaatgc cttggccttt atatggaaat gtggaactga gtggatatgc
     2041 tgtttttgtc tgttaaacag agaagctggc tgttatccac tgagaagcga acgaaacagt
     2101 cgggaaaatc tcccattatc gtagagatcc gcattattaa tctcaggagc ctgtgtagcg
     2161 tttataggaa gtagtgttct gtcatgatgc ctgcaagcgg taacgaaaac gatttgaata
     2221 tgccttcagg aacaatagaa atcttcgtgc ggtgttacgt tgaagtggag cggattatgt
     2281 cagcaatgga cagaacaacc taatgaacac agaaccatga tgtggtctgt ccttttacag
     2341 ccagtagtgc tcgccgcagt cgagcgacag ggcgaagccc tcgagtgagc gaggaagcac
     2401 cagggaacag cacttatata ttctgcttac acacgatgcc tgaaaaaact tcccttgggg
     2461 ttatccactt atccacgggg atatttttat aattattttt tttatagttt ttagatcttc
     2521 ttttttagag cgccttgtag gcctttatcc atgctggttc tagagaaggt gttgtgacaa
     2581 attgcccttt cagtgtgaca aatcaccctc aaatgacagt cctgtctgtg acaaattgcc
     2641 cttaaccctg tgacaaattg ccctcagaag aagctgtttt ttcacaaagt tatccctgct
     2701 tattgactct tttttattta gtgtgacaat ctaaaaactt gtcacacttc acatggatct
     2761 gtcatggcgg aaacagcggt tatcaatcac aagaaacgta aaaatagccc gcgaatcgtc
     2821 cagtcaaacg acctcactga ggcggcatat agtctctccc gggatcaaaa acgtatgctg
     2881 tatctgttcg ttgaccagat cagaaaatct gatggcaccc tacaggaaca tgacggtatc
     2941 tgcgagatcc atgttgctaa atatgctgaa atattcggat tgacctctgc ggaagccagt
     3001 aaggatatac ggcaggcatt gaagagtttc gcggggaagg aagtggtttt ttatcgccct
     3061 gaagaggatg ccggcgatga aaaaggctat gaatcttttc cttggtttat caaacgtgcg
     3121 cacagtccat ccagagggct ttacagtgta catatcaacc catatctcat tcccttcttt
     3181 atcgggttac agaaccggtt tacgcagttt cggcttagtg aaacaaaaga aatcaccaat
     3241 ccgtatgcca tgcgtttata cgaatccctg tgtcagtatc gtaagccgga tggctcaggc
     3301 atcgtctctc tgaaaatcga ctggatcata gagcgttacc agctgcctca aagttaccag
     3361 cgtatgcctg acttccgccg ccgcttcctg caggtctgtg ttaatgagat caacagcaga
     3421 actccaatgc gcctctcata cattgagaaa aagaaaggcc gccagacgac tcatatcgta
     3481 ttttccttcc gcgatatcac ttccatgacg acaggatagt ctgagggtta tctgtcacag
     3541 atttgagggt ggttcgtcac atttgttctg acctactgag ggtaatttgt cacagttttg
     3601 ctgtttcctt cagcctgcat ggattttctc atactttttg aactgtaatt tttaaggaag
     3661 ccaaatttga gggcagtttg tcacagttga tttccttctc tttcccttcg tcatgtgacc
     3721 tgatatcggg ggttagttcg tcatcattga tgagggttga ttatcacagt ttattactct
     3781 gaattggcta tccgcgtgtg tacctctacc tggagttttt cccacggtgg atatttcttc
     3841 ttgcgctgag cgtaagagct atctgacaga acagttcttc tttgcttcct cgccagttcg
     3901 ctcgctatgc tcggttacac ggctgcggcg agcgctagtg ataataagtg actgaggtat
     3961 gtgctcttct tatctccttt tgtagtgttg ctcttatttt aaacaacttt gcggtttttt
     4021 gatgactttg cgattttgtt gttgctttgc agtaaattgc aagatttaat aaaaaaacgc
     4081 aaagcaatga ttaaaggatg ttcagaatga aactcatgga aacacttaac cagtgcataa
     4141 acgctggtca tgaaatgacg aaggctatcg ccattgcaca gtttaatgat gacagcccgg
     4201 aagcgaggaa aataacccgg cgctggagaa taggtgaagc agcggattta gttggggttt
     4261 cttctcaggc tatcagagat gccgagaaag cagggcgact accgcacccg gatatggaaa
     4321 ttcgaggacg ggttgagcaa cgtgttggtt atacaattga acaaattaat catatgcgtg
     4381 atgtgtttgg tacgcgattg cgacgtgctg aagacgtatt tccaccggtg atcggggttg
     4441 ctgcccataa aggtggcgtt tacaaaacct cagtttctgt tcatcttgct caggatctgg
     4501 ctctgaaggg gctacgtgtt ttgctcgtgg aaggtaacga cccccaggga acagcctcaa
     4561 tgtatcacgg atgggtacca gatcttcata ttcatgcaga agacactctc ctgcctttct
     4621 atcttgggga aaaggacgat gtcacttatg caataaagcc cacttgctgg ccggggcttg
     4681 acattattcc ttcctgtctg gctctgcacc gtattgaaac tgagttaatg ggcaaatttg
     4741 atgaaggtaa actgcccacc gatccacacc tgatgctccg actggccatt gaaactgttg
     4801 ctcatgacta tgatgtcata gttattgaca gcgcgcctaa cctgggtatc ggcacgatta
     4861 atgtcgtatg tgctgctgat gtgctgattg ttcccacgcc tgctgagttg tttgactaca
     4921 cctccgcact gcagtttttc gatatgcttc gtgatctgct caagaacgtt gatcttaaag
     4981 ggttcgagcc tgatgtacgt attttgctta ccaaatacag caatagtaat ggctctcagt
     5041 ccccgtggat ggaggagcaa attcgggatg cctggggaag catggttcta aaaaatgttg
     5101 tacgtgaaac ggatgaagtt ggtaaaggtc agatccggat gagaactgtt tttgaacagg
     5161 ccattgatca acgctcttca actggtgcct ggagaaatgc tctttctatt tgggaacctg
     5221 tctgcaatga aattttcgat cgtctgatta aaccacgctg ggagattaga taatgaagcg
     5281 tgcgcctgtt attccaaaac atacgctcaa tactcaaccg gttgaagata cttcgttatc
     5341 gacaccagct gccccgatgg tggattcgtt aattgcgcgc gtaggagtaa tggctcgcgg
     5401 taatgccatt actttgcctg tatgtggtcg ggatgtgaag tttactcttg aagtgctccg
     5461 gggtgatagt gttgagaaga cctctcgggt atggtcaggt aatgaacgtg accaggagct
     5521 gcttactgag gacgcactgg atgatctcat cccttctttt ctactgactg gtcaacagac
     5581 accggcgttc ggtcgaagag tatctggtgt catagaaatt gccgatggga gtcgccgtcg
     5641 taaagctgct gcacttaccg aaagtgatta tcgtgttctg gttggcgagc tggatgatga
     5701 gcagatggct gcattatcca gattgggtaa cgattatcgc ccaacaagtg cttatgaacg
     5761 tggtcagcgt tatgcaagcc gattgcagaa tgaatttgct ggaaatattt ctgcgctggc
     5821 tgatgcggaa aatatttcac gtaagattat tacccgctgt atcaacaccg ccaaattgcc
     5881 taaatcagtt gttgctcttt tttctcaccc cggtgaacta tctgcccggt caggtgatgc
     5941 acttcaaaaa gcctttacag ataaagagga attacttaag cagcaggcat ctaaccttca
     6001 tgagcagaaa aaagctgggg tgatatttga agctgaagaa gttatcactc ttttaacttc
     6061 tgtgcttaaa acgtcatctg catcaagaac tagtttaagc tcacgacatc agtttgctcc
     6121 tggagcgaca gtattgtata agggcgataa aatggtgctt aacctggaca ggtctcgtgt
     6181 tccaactgag tgtatagaga aaattgaggc cattcttaag gaacttgaaa agccagcacc
     6241 ctgatgcgac cacgttttag tctacgttta tctgtcttta cttaatgtcc tttgttacag
     6301 gccagaaagc ataactggcc tgaatattct ctctgggccc actgttccac ttgtatcgtc
     6361 ggtctgataa tcagactggg accacggtcc cactcgtatc gtcggtctga ttattagtct
     6421 gggaccacgg tcccactcgt atcgtcggtc tgattattag tctgggacca cggtcccact
     6481 cgtatcgtcg gtctgataat cagactggga ccacggtccc actcgtatcg tcggtctgat
     6541 tattagtctg ggaccatggt cccactcgta tcgtcggtct gattattagt ctgggaccac
     6601 ggtcccactc gtatcgtcgg tctgattatt agtctggaac cacggtccca ctcgtatcgt
     6661 cggtctgatt attagtctgg gaccacggtc ccactcgtat cgtcggtctg attattagtc
     6721 tgggaccacg atcccactcg tgttgtcggt ctgattatcg gtctgggacc acggtcccac
     6781 ttgtattgtc gatcagacta tcagcgtgag actacgattc catcaatgcc tgtcaagggc
     6841 aagtattgac atgtcgtcgt aacctgtaga acggagtaac ctcggtgtgc ggttgtatgc
     6901 ctgctgtgga ttgctgctgt gtcctgctta tccacaacat tttgcgcacg gttatgtgga
     6961 caaaatacct ggttacccag gccgtgccgg cacgttaacc gggctgcatc cgatgcaagt
     7021 gtgtcgctgt cgacgagctc gcgagctcgg acatgaggtt gccccgtatt cagtgtcgct
     7081 gatttgtatt gtctgaagtt gtttttacgt taagttgatg cagatcaatt aatacgatac
     7141 ctgcgtcata attgattatt tgacgtggtt tgatggcctc cacgcacgtt gtgatatgta
     7201 gatgataatc attatcactt tacgggtcct ttccggtgat ccgacaggtt acggggcggc
     7261 gacctcgcgg gttttcgcta tttatgaaaa ttttccggtt taaggcgttt ccgttcttct
     7321 tcgtcataac ttaatgtttt tatttaaaat accctctgaa aagaaaggaa acgacaggtg
     7381 ctgaaagcga gctttttggc ctctgtcgtt tcctttctct gtttttgtcc gtggaatgaa
     7441 caatggaagt ccgagctcat cgctaataac ttcgtatagc atacattata cgaagttata
     7501 ttcgat
//
