LOCUS       Exported                5655 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000519 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5655)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 2, 2021 from SnapGene 5.3.1
            https://www.snapgene.com
COMMENT     This file was created using tools provided by Twist Bioscience
FEATURES             Location/Qualifiers
     source          1..5655
                     /organism="BBF10K_000519 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1161
                     /codon_start=1
                     /product="platelet-derived growth factor D transcription 
                     variant 1"
                     /label=PDGF-DDtv1
                     /translation="MHRLIFVYTLICANFCSCRDTSATPQSASIKALRNANLRRDESNH
                     LTDLYRRDETIQVKGNGYVQSPRFPNSYPRNLLLTWRLHSQENTRIQLVFDNQFGLEEA
                     ENDICRYDFVEVEDISETSTIIRGRWCGHKEVPPRIKSRTNQIKITFKSDDYFVAKPGF
                     KIYYSLLEDFQPAAASETNWESVTSSISGVSYNSPSVTDPTLIADALDKKIAEFDTVED
                     LLKYFNPESWQEDLENMYLDTPRYRGRSYHDRKSKVDLDRLNDDAKRYSCTPRNYSVNI
                     REELKLANVVFFPRCLLVQRCGGNCGCGTVNWRSCTCNSGKTVKKYHEVLQFEPGHIKR
                     RGRAKTMALVDIQLDHHERCDCICSSRPPR"
     primer_bind     complement(1162..1186)
                     /label=Gibson-rev
     terminator      1305..1352
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1357..1373)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1532..1782
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1785..2540)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2631..2850
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3270..3291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3306..3336
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3344..3360
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     3368..3384
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3457..4302
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4410..4561
                     /label=nnpp promoters
     CDS             4601..5416
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     terminator      complement(5526..5569)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5587..5603
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5604..5635
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgcacaga
       61 ctgatcttcg tgtacaccct gatctgcgcc aacttctgca gctgcagaga caccagcgcc
      121 accccccaga gcgccagcat caaggccctg agaaacgcca acctgagaag agacgagagc
      181 aaccacctga ccgacctgta cagaagagac gagaccatcc aggtgaaggg caacggctac
      241 gtgcagagcc ccagattccc caacagctac cccagaaacc tgctgctgac ctggagactg
      301 cacagccagg agaacaccag aatccagctg gtgttcgaca accagttcgg cctggaggag
      361 gccgagaacg acatctgcag atacgacttc gtggaggtgg aggacatcag cgagaccagc
      421 accatcatca gaggcagatg gtgcggccac aaggaggtgc cccccagaat caagagcaga
      481 accaaccaga tcaagatcac cttcaagagc gacgactact tcgtggccaa gcccggcttc
      541 aagatctact acagcctgct ggaggacttc cagcccgccg ccgccagcga gaccaactgg
      601 gagagcgtga ccagcagcat cagcggcgtg agctacaaca gccccagcgt gaccgacccc
      661 accctgatcg ccgacgccct ggacaagaag atcgccgagt tcgacaccgt ggaggacctg
      721 ctgaagtact tcaaccccga gagctggcag gaggacctgg agaacatgta cctggacacc
      781 cccagataca gaggcagaag ctaccacgac agaaagagca aggtggacct ggacagactg
      841 aacgacgacg ccaagagata cagctgcacc cccagaaact acagcgtgaa catcagagag
      901 gagctgaagc tggccaacgt ggtgttcttc cccagatgcc tgctggtgca gagatgcggc
      961 ggcaactgcg gctgcggcac cgtgaactgg agaagctgca cctgcaacag cggcaagacc
     1021 gtgaagaagt accacgaggt gctgcagttc gagcccggcc acatcaagag aagaggcaga
     1081 gccaagacca tggccctggt ggacatccag ctggaccacc acgagagatg cgactgcatc
     1141 tgcagcagca gaccccccag aggatccgcc accaacttca gcctgctaga ttgttactcc
     1201 gacggacaga gggaaaggca ggagcggcta gggataacag ggtaatgagt tccttaaacg
     1261 ttatgttcgc tgaggtgtca atcgtcggag ccgctgagca ataactagca taaccccttg
     1321 gggcctctaa acgggtcttg aggggttttt tgcatggtca tagctgtttc ctgagagctt
     1381 ggcaggtgtc gcgtatgcat cgatcacgtg tccacgttct ttaatagtgg actcttgttc
     1441 caaactggaa caacactcgg atcgatccgg cgcgcaccgt gggaaaaact ccaggtagag
     1501 gtacacacgc ggatagccaa ttcagagtaa taaactgtga taatcaaccc tcatcaatga
     1561 tgacgaacta acccccgata tcaggtcaca tgacgaaggg aaagagaagg aaatcaactg
     1621 tgacaaactg ccctcaaatt tggcttcctt aaaaattaca gttcaaaaag tatgagaaaa
     1681 tccatgcagg ctgaaggaaa cagcaaaact gtgacaaatt accctcagta ggtcagaaca
     1741 aatgtgacga accaccctca aatctgtgac agataaccct cagactatcc tgtcgtcatg
     1801 gaagtgatat cgcggaagga aaatacgata tgagtcgtct ggcggccttt ctttttctca
     1861 atgtatgaga ggcgcattgg agttctgctg ttgatctcat taacacagac ttgcaggaag
     1921 cggcggcgga agtcaggcat acgctggtaa ctttgaggca gctggtaacg ctctatgatc
     1981 cagtcgattt tcagagacac gatgcctgag ccatccggct tacgatactg acacagggat
     2041 tcgtataaac gcatggcata cggattggtg atttcttttg tttcactaag ccgaaactgc
     2101 gtaaaccggt tctgtaaccc gataaagaag ggaatgagat atgggttgat atgtacactg
     2161 taaagccctc tggatggact gtgcgcacgt ttgataaacc aaggaaaaga ttcatagcct
     2221 ttttcatcac cggcatcctc ttcagggcga taaaaaacca cttccttccc cgcgaaactc
     2281 ttcaatgcct gccgtatatc cttactggct tccgcagagg tcaatccgaa tatttcagca
     2341 tatttagcaa catggatctc gcagataccg tcatgttcct gtagggtgcc atcagatttt
     2401 ctgatctggt caacgaacag atacagcata cgtttttgat cccgggagag actatatgcc
     2461 gcctcagtga ggtcgtttga ctggacgatt cgcgggctat ttttacgttt cttgtgattg
     2521 ataaccgctg tttccgccat gacagatcca tgtgaagtgt gacaagtttt tagattgtca
     2581 cactaaataa aaaagagtca ataagcaggg ataactttgt gaaaaaacag cttcttctga
     2641 gggcaatttg tcacagggtt aagggcaatt tgtcacagac aggactgtca tttgagggtg
     2701 atttgtcaca ctgaaagggc aatttgtcac aacaccttct gtagaaccag catggataaa
     2761 ggcctacaag gcgctctaaa aaagaagatc taaaaactat aaaaaaaata attataaaaa
     2821 tatccccgtg gataagtgga taaccccaag ggaagttttt tcaggcatcg tgtgtaagca
     2881 gaatatataa gtgctgttcc ctggtgcttc ctcgctcact cgaaattccc ggggatagct
     2941 ttatgcttgt aaaccgtttt gtgaaaaaat ttttaaaata aaaaagggga cctctagggt
     3001 ccccaattaa ttagtaatat aatctattaa aggtcattca aaaggtcatc caccggatca
     3061 attcccctgc tcgcgcaggc tgggtgccaa gctctcgggt aacatcaagg cccgatcctt
     3121 ggagcccttg ccctcccgca cgatgatcgt gccgtgatcg aaatccagat ccttgacccg
     3181 cagttgcaaa ccctcactga tccgattcat taatgcagct ggcacgacag gtttcccgac
     3241 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     3301 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     3361 tttcacacag gaaacagcta tgaccatgat tacgccaagc gcggtacctg tcgcggcaac
     3421 gcgctaacag acgtagtaag aaccaccagc attgtaatgc tggctaaagt cactttcctg
     3481 agctgtataa cgatgagcga ttttactttt tctggctatg aattggcctg ctttgtaaca
     3541 cactccggtc tatcccgtag cgccgggcat atcctgtcgc aatgtgcaaa tctcgcggca
     3601 acaaccagtg aatacttcat tcacaagcct caccgcctga tcgcggcaga aactggttat
     3661 agccaatcaa ccgtcgttcg tgcattccgt gaagctgtaa acaaaggtat cctgtctgta
     3721 gagattgtta tcggcgatca ccgtgaacgt cgcgctaacc tgtaccggtt tacaccatcc
     3781 tttttggcct tcgcacaaca agccaaaaat gcgctgatag aaagcaaatt aaagatctct
     3841 tcagcggcaa ccaaggttaa agctgttctc gctaagacat tggctttatt taatttttta
     3901 tccacacccc catgtcaaaa tgataccccc tccccctgtc aggatgacgt ggcaataaag
     3961 aataagaagt cacaagttaa aaaaacaaaa agatcagttt ccggcggtgc cggaacaacc
     4021 agcctcaaaa aattgacttc atggatcgct aaggcaaaag caaaggctga caatctgcgg
     4081 ttatccaaaa aacgcactca aaaacatgag ttcaagcaga aagtagaggc ggctgcgcgg
     4141 aaatatgctt acctgaagaa caagcgttcg cctgatattg gcgggatatc aaacttcgat
     4201 aacctaccgc attgcatgac ggtaaacgaa gctcttaatg cggttttagc caaaaataaa
     4261 gataacgaac aatggggtat accggcagga ttcagagggt aatgaattgc tctaattata
     4321 accatgcata ctttcaacac ctctagtttg ccatgaggca aactcatagg tgtcctggta
     4381 agaggacact gttgccaaaa ctggacgccc cattattgca attaataaac aactaacgga
     4441 caattctacc taacaataag tggagttgcg gccggccgcg atccgctagc aaagccacgt
     4501 tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat
     4561 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga
     4621 aacgtcttgc tcgcgcccgc gattaaattc caacatggat gctgatttat atgggtataa
     4681 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc
     4741 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
     4801 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
     4861 tatccgtact cctgatgatg catggttact caccactgcg atccccggga aaacagcatt
     4921 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
     4981 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
     5041 tcgcctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
     5101 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aactgttgcc
     5161 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
     5221 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     5281 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     5341 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     5401 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcgag ctctgccacc
     5461 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     5521 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct gcacctgcac
     5581 cagtcagtaa aacgacggcc agtagtcaaa agcctccgac cggaggcttt tgacttggtt
     5641 caggtggagt gggag
//
