LOCUS       Exported                5637 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000520 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5637)
  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..5637
                     /organism="BBF10K_000520 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1143
                     /codon_start=1
                     /product="platelet-derived growth factor D transcription 
                     variant 2"
                     /label=PDGF-DDtv2
                     /translation="MHRLIFVYTLICANFCSCRDTSATPQSASIKALRNANLRRDDLYR
                     RDETIQVKGNGYVQSPRFPNSYPRNLLLTWRLHSQENTRIQLVFDNQFGLEEAENDICR
                     YDFVEVEDISETSTIIRGRWCGHKEVPPRIKSRTNQIKITFKSDDYFVAKPGFKIYYSL
                     LEDFQPAAASETNWESVTSSISGVSYNSPSVTDPTLIADALDKKIAEFDTVEDLLKYFN
                     PESWQEDLENMYLDTPRYRGRSYHDRKSKVDLDRLNDDAKRYSCTPRNYSVNIREELKL
                     ANVVFFPRCLLVQRCGGNCGCGTVNWRSCTCNSGKTVKKYHEVLQFEPGHIKRRGRAKT
                     MALVDIQLDHHERCDCICSSRPPR"
     primer_bind     complement(1144..1168)
                     /label=Gibson-rev
     terminator      1287..1334
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1339..1355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1514..1764
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1767..2522)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2613..2832
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3252..3273
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3288..3318
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3326..3342
                     /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     3350..3366
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3439..4284
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4392..4543
                     /label=nnpp promoters
     CDS             4583..5398
                     /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(5508..5551)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5569..5585
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5586..5617
                     /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 agacgacctg
      181 tacagaagag acgagaccat ccaggtgaag ggcaacggct acgtgcagag ccccagattc
      241 cccaacagct accccagaaa cctgctgctg acctggagac tgcacagcca ggagaacacc
      301 agaatccagc tggtgttcga caaccagttc ggcctggagg aggccgagaa cgacatctgc
      361 agatacgact tcgtggaggt ggaggacatc agcgagacca gcaccatcat cagaggcaga
      421 tggtgcggcc acaaggaggt gccccccaga atcaagagca gaaccaacca gatcaagatc
      481 accttcaaga gcgacgacta cttcgtggcc aagcccggct tcaagatcta ctacagcctg
      541 ctggaggact tccagcccgc cgccgccagc gagaccaact gggagagcgt gaccagcagc
      601 atcagcggcg tgagctacaa cagccccagc gtgaccgacc ccaccctgat cgccgacgcc
      661 ctggacaaga agatcgccga gttcgacacc gtggaggacc tgctgaagta cttcaacccc
      721 gagagctggc aggaggacct ggagaacatg tacctggaca cccccagata cagaggcaga
      781 agctaccacg acagaaagag caaggtggac ctggacagac tgaacgacga cgccaagaga
      841 tacagctgca cccccagaaa ctacagcgtg aacatcagag aggagctgaa gctggccaac
      901 gtggtgttct tccccagatg cctgctggtg cagagatgcg gcggcaactg cggctgcggc
      961 accgtgaact ggagaagctg cacctgcaac agcggcaaga ccgtgaagaa gtaccacgag
     1021 gtgctgcagt tcgagcccgg ccacatcaag agaagaggca gagccaagac catggccctg
     1081 gtggacatcc agctggacca ccacgagaga tgcgactgca tctgcagcag cagacccccc
     1141 agaggatccg ccaccaactt cagcctgcta gatttccgac gagattccct agccgtttat
     1201 aggctgatta tagggataac agggtaatag accgtgaggt aatgtctggc gctgaggtgt
     1261 caatcgtcgg agccgctgag caataactag cataacccct tggggcctct aaacgggtct
     1321 tgaggggttt tttgcatggt catagctgtt tcctgagagc ttggcaggtg tcgcgtatgc
     1381 atcgatcacg tgtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1441 ggatcgatcc ggcgcgcacc gtgggaaaaa ctccaggtag aggtacacac gcggatagcc
     1501 aattcagagt aataaactgt gataatcaac cctcatcaat gatgacgaac taacccccga
     1561 tatcaggtca catgacgaag ggaaagagaa ggaaatcaac tgtgacaaac tgccctcaaa
     1621 tttggcttcc ttaaaaatta cagttcaaaa agtatgagaa aatccatgca ggctgaagga
     1681 aacagcaaaa ctgtgacaaa ttaccctcag taggtcagaa caaatgtgac gaaccaccct
     1741 caaatctgtg acagataacc ctcagactat cctgtcgtca tggaagtgat atcgcggaag
     1801 gaaaatacga tatgagtcgt ctggcggcct ttctttttct caatgtatga gaggcgcatt
     1861 ggagttctgc tgttgatctc attaacacag acttgcagga agcggcggcg gaagtcaggc
     1921 atacgctggt aactttgagg cagctggtaa cgctctatga tccagtcgat tttcagagac
     1981 acgatgcctg agccatccgg cttacgatac tgacacaggg attcgtataa acgcatggca
     2041 tacggattgg tgatttcttt tgtttcacta agccgaaact gcgtaaaccg gttctgtaac
     2101 ccgataaaga agggaatgag atatgggttg atatgtacac tgtaaagccc tctggatgga
     2161 ctgtgcgcac gtttgataaa ccaaggaaaa gattcatagc ctttttcatc accggcatcc
     2221 tcttcagggc gataaaaaac cacttccttc cccgcgaaac tcttcaatgc ctgccgtata
     2281 tccttactgg cttccgcaga ggtcaatccg aatatttcag catatttagc aacatggatc
     2341 tcgcagatac cgtcatgttc ctgtagggtg ccatcagatt ttctgatctg gtcaacgaac
     2401 agatacagca tacgtttttg atcccgggag agactatatg ccgcctcagt gaggtcgttt
     2461 gactggacga ttcgcgggct atttttacgt ttcttgtgat tgataaccgc tgtttccgcc
     2521 atgacagatc catgtgaagt gtgacaagtt tttagattgt cacactaaat aaaaaagagt
     2581 caataagcag ggataacttt gtgaaaaaac agcttcttct gagggcaatt tgtcacaggg
     2641 ttaagggcaa tttgtcacag acaggactgt catttgaggg tgatttgtca cactgaaagg
     2701 gcaatttgtc acaacacctt ctgtagaacc agcatggata aaggcctaca aggcgctcta
     2761 aaaaagaaga tctaaaaact ataaaaaaaa taattataaa aatatccccg tggataagtg
     2821 gataacccca agggaagttt tttcaggcat cgtgtgtaag cagaatatat aagtgctgtt
     2881 ccctggtgct tcctcgctca ctcgaaattc ccggggatag ctttatgctt gtaaaccgtt
     2941 ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt aattagtaat
     3001 ataatctatt aaaggtcatt caaaaggtca tccaccggat caattcccct gctcgcgcag
     3061 gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc ttggagccct tgccctcccg
     3121 cacgatgatc gtgccgtgat cgaaatccag atccttgacc cgcagttgca aaccctcact
     3181 gatccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     3241 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     3301 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     3361 tatgaccatg attacgccaa gcgcggtacc tgtcgcggca acgcgctaac agacgtagta
     3421 agaaccacca gcattgtaat gctggctaaa gtcactttcc tgagctgtat aacgatgagc
     3481 gattttactt tttctggcta tgaattggcc tgctttgtaa cacactccgg tctatcccgt
     3541 agcgccgggc atatcctgtc gcaatgtgca aatctcgcgg caacaaccag tgaatacttc
     3601 attcacaagc ctcaccgcct gatcgcggca gaaactggtt atagccaatc aaccgtcgtt
     3661 cgtgcattcc gtgaagctgt aaacaaaggt atcctgtctg tagagattgt tatcggcgat
     3721 caccgtgaac gtcgcgctaa cctgtaccgg tttacaccat cctttttggc cttcgcacaa
     3781 caagccaaaa atgcgctgat agaaagcaaa ttaaagatct cttcagcggc aaccaaggtt
     3841 aaagctgttc tcgctaagac attggcttta tttaattttt tatccacacc cccatgtcaa
     3901 aatgataccc cctccccctg tcaggatgac gtggcaataa agaataagaa gtcacaagtt
     3961 aaaaaaacaa aaagatcagt ttccggcggt gccggaacaa ccagcctcaa aaaattgact
     4021 tcatggatcg ctaaggcaaa agcaaaggct gacaatctgc ggttatccaa aaaacgcact
     4081 caaaaacatg agttcaagca gaaagtagag gcggctgcgc ggaaatatgc ttacctgaag
     4141 aacaagcgtt cgcctgatat tggcgggata tcaaacttcg ataacctacc gcattgcatg
     4201 acggtaaacg aagctcttaa tgcggtttta gccaaaaata aagataacga acaatggggt
     4261 ataccggcag gattcagagg gtaatgaatt gctctaatta taaccatgca tactttcaac
     4321 acctctagtt tgccatgagg caaactcata ggtgtcctgg taagaggaca ctgttgccaa
     4381 aactggacgc cccattattg caattaataa acaactaacg gacaattcta cctaacaata
     4441 agtggagttg cggccggccg cgatccgcta gcaaagccac gttgtgtctc aaaatctctg
     4501 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
     4561 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtctt gctcgcgccc
     4621 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     4681 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     4741 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     4801 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     4861 tgcatggtta ctcaccactg cgatccccgg gaaaacagca ttccaggtat tagaagaata
     4921 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     4981 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgcctcg ctcaggcgca
     5041 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     5101 gcctgttgaa caagtctgga aagaaatgca taaactgttg ccattctcac cggattcagt
     5161 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     5221 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     5281 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     5341 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     5401 agaattggtt aattggttgt aacactggcg agctctgcca cctgacgtct aagaaaccat
     5461 tattatcatg acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa
     5521 aaaatcctta gctttcgcta aggatgattt ctgcacctgc accagtcagt aaaacgacgg
     5581 ccagtagtca aaagcctccg accggaggct tttgacttgg ttcaggtgga gtgggag
//
