LOCUS       Exported                5148 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000521 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5148)
  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..5148
                     /organism="BBF10K_000521 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..654
                     /codon_start=1
                     /product="HSPG syndecan 2"
                     /label=SDC2
                     /translation="MRRAWILLTLGLVACVSAESRAELTSDKDMYLDNSSIEEASGVYP
                     IDDDDYASASGSGADEDVESPELTTSRPLPKILLTSAAPKVETTTLNIQNKIPAQTKSP
                     EETDKEKVHLSDSERKMDPAEEDTNVYTEKHSDSLFKRTEVLAAVIAGGVIGFLFAIFL
                     ILLLVYRMRKKDEGSYDLGERKPSSAAYQKAPTKEFYA"
     primer_bind     complement(655..679)
                     /label=Gibson-rev
     terminator      798..845
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(850..866)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1025..1275
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1278..2033)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2124..2343
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2763..2784
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2799..2829
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2837..2853
                     /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     2861..2877
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2950..3795
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    3903..4054
                     /label=nnpp promoters
     CDS             4094..4909
                     /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(5019..5062)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5080..5096
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5097..5128
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgagaaga
       61 gcctggatac tgctgaccct gggcctggtg gcctgcgtga gcgccgagag cagagccgag
      121 ctgaccagcg acaaggacat gtacctggac aacagcagca tcgaggaggc cagcggcgtg
      181 taccccatcg acgacgacga ctacgccagc gccagcggca gcggcgccga cgaggacgtg
      241 gagagccccg agctgaccac cagcagaccc ctgcccaaga tcctgctgac cagcgccgcc
      301 cccaaggtgg agaccaccac cctgaacatc cagaacaaga tccccgccca gaccaagagc
      361 cccgaggaga ccgacaagga gaaggtgcac ctgagcgaca gcgagagaaa gatggacccc
      421 gccgaggagg acaccaacgt gtacaccgag aagcacagcg acagcctgtt caagagaacc
      481 gaggtgctgg ccgccgtgat cgccggcggc gtgatcggct tcctgttcgc catcttcctg
      541 atcctgctgc tggtgtacag aatgagaaag aaggacgagg gcagctacga cctgggcgag
      601 agaaagccca gcagcgccgc ctaccagaag gcccccacca aggagttcta cgccggatcc
      661 gccaccaact tcagcctgct agcatccgtc taaatctcgg ttagtgcact ctcaccgacg
      721 atagggataa cagggtaatg aagtgtgggc tggaggtaag cgctgaggtg tcaatcgtcg
      781 gagccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      841 ttttgcatgg tcatagctgt ttcctgagag cttggcaggt gtcgcgtatg catcgatcac
      901 gtgtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact cggatcgatc
      961 cggcgcgcac cgtgggaaaa actccaggta gaggtacaca cgcggatagc caattcagag
     1021 taataaactg tgataatcaa ccctcatcaa tgatgacgaa ctaacccccg atatcaggtc
     1081 acatgacgaa gggaaagaga aggaaatcaa ctgtgacaaa ctgccctcaa atttggcttc
     1141 cttaaaaatt acagttcaaa aagtatgaga aaatccatgc aggctgaagg aaacagcaaa
     1201 actgtgacaa attaccctca gtaggtcaga acaaatgtga cgaaccaccc tcaaatctgt
     1261 gacagataac cctcagacta tcctgtcgtc atggaagtga tatcgcggaa ggaaaatacg
     1321 atatgagtcg tctggcggcc tttctttttc tcaatgtatg agaggcgcat tggagttctg
     1381 ctgttgatct cattaacaca gacttgcagg aagcggcggc ggaagtcagg catacgctgg
     1441 taactttgag gcagctggta acgctctatg atccagtcga ttttcagaga cacgatgcct
     1501 gagccatccg gcttacgata ctgacacagg gattcgtata aacgcatggc atacggattg
     1561 gtgatttctt ttgtttcact aagccgaaac tgcgtaaacc ggttctgtaa cccgataaag
     1621 aagggaatga gatatgggtt gatatgtaca ctgtaaagcc ctctggatgg actgtgcgca
     1681 cgtttgataa accaaggaaa agattcatag cctttttcat caccggcatc ctcttcaggg
     1741 cgataaaaaa ccacttcctt ccccgcgaaa ctcttcaatg cctgccgtat atccttactg
     1801 gcttccgcag aggtcaatcc gaatatttca gcatatttag caacatggat ctcgcagata
     1861 ccgtcatgtt cctgtagggt gccatcagat tttctgatct ggtcaacgaa cagatacagc
     1921 atacgttttt gatcccggga gagactatat gccgcctcag tgaggtcgtt tgactggacg
     1981 attcgcgggc tatttttacg tttcttgtga ttgataaccg ctgtttccgc catgacagat
     2041 ccatgtgaag tgtgacaagt ttttagattg tcacactaaa taaaaaagag tcaataagca
     2101 gggataactt tgtgaaaaaa cagcttcttc tgagggcaat ttgtcacagg gttaagggca
     2161 atttgtcaca gacaggactg tcatttgagg gtgatttgtc acactgaaag ggcaatttgt
     2221 cacaacacct tctgtagaac cagcatggat aaaggcctac aaggcgctct aaaaaagaag
     2281 atctaaaaac tataaaaaaa ataattataa aaatatcccc gtggataagt ggataacccc
     2341 aagggaagtt ttttcaggca tcgtgtgtaa gcagaatata taagtgctgt tccctggtgc
     2401 ttcctcgctc actcgaaatt cccggggata gctttatgct tgtaaaccgt tttgtgaaaa
     2461 aatttttaaa ataaaaaagg ggacctctag ggtccccaat taattagtaa tataatctat
     2521 taaaggtcat tcaaaaggtc atccaccgga tcaattcccc tgctcgcgca ggctgggtgc
     2581 caagctctcg ggtaacatca aggcccgatc cttggagccc ttgccctccc gcacgatgat
     2641 cgtgccgtga tcgaaatcca gatccttgac ccgcagttgc aaaccctcac tgatccgatt
     2701 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2761 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     2821 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     2881 gattacgcca agcgcggtac ctgtcgcggc aacgcgctaa cagacgtagt aagaaccacc
     2941 agcattgtaa tgctggctaa agtcactttc ctgagctgta taacgatgag cgattttact
     3001 ttttctggct atgaattggc ctgctttgta acacactccg gtctatcccg tagcgccggg
     3061 catatcctgt cgcaatgtgc aaatctcgcg gcaacaacca gtgaatactt cattcacaag
     3121 cctcaccgcc tgatcgcggc agaaactggt tatagccaat caaccgtcgt tcgtgcattc
     3181 cgtgaagctg taaacaaagg tatcctgtct gtagagattg ttatcggcga tcaccgtgaa
     3241 cgtcgcgcta acctgtaccg gtttacacca tcctttttgg ccttcgcaca acaagccaaa
     3301 aatgcgctga tagaaagcaa attaaagatc tcttcagcgg caaccaaggt taaagctgtt
     3361 ctcgctaaga cattggcttt atttaatttt ttatccacac ccccatgtca aaatgatacc
     3421 ccctccccct gtcaggatga cgtggcaata aagaataaga agtcacaagt taaaaaaaca
     3481 aaaagatcag tttccggcgg tgccggaaca accagcctca aaaaattgac ttcatggatc
     3541 gctaaggcaa aagcaaaggc tgacaatctg cggttatcca aaaaacgcac tcaaaaacat
     3601 gagttcaagc agaaagtaga ggcggctgcg cggaaatatg cttacctgaa gaacaagcgt
     3661 tcgcctgata ttggcgggat atcaaacttc gataacctac cgcattgcat gacggtaaac
     3721 gaagctctta atgcggtttt agccaaaaat aaagataacg aacaatgggg tataccggca
     3781 ggattcagag ggtaatgaat tgctctaatt ataaccatgc atactttcaa cacctctagt
     3841 ttgccatgag gcaaactcat aggtgtcctg gtaagaggac actgttgcca aaactggacg
     3901 ccccattatt gcaattaata aacaactaac ggacaattct acctaacaat aagtggagtt
     3961 gcggccggcc gcgatccgct agcaaagcca cgttgtgtct caaaatctct gatgttacat
     4021 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     4081 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctcgcgcc cgcgattaaa
     4141 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     4201 aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     4261 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     4321 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     4381 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     4441 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     4501 ttgtaattgt ccttttaaca gcgatcgcgt atttcgcctc gctcaggcgc aatcacgaat
     4561 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     4621 acaagtctgg aaagaaatgc ataaactgtt gccattctca ccggattcag tcgtcactca
     4681 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     4741 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     4801 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     4861 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     4921 taattggttg taacactggc gagctctgcc acctgacgtc taagaaacca ttattatcat
     4981 gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt
     5041 agctttcgct aaggatgatt tctgcacctg caccagtcag taaaacgacg gccagtagtc
     5101 aaaagcctcc gaccggaggc ttttgacttg gttcaggtgg agtgggag
//
