LOCUS       Exported                4947 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000539 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4947)
  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..4947
                     /organism="BBF10K_000539 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..453
                     /codon_start=1
                     /product="inhibitor of DNA binding 2"
                     /label=ID2
                     /translation="MKAFSPVRSVRKNSLSDHSLGISRSKTPVDDPMSLLYNMNDCYSK
                     LKELVPSIPQNKKVSKMEILQHVIDYILDLQIALDSHPTIVSLHHQRPGQNQASRTPLT
                     TLNTDISILSLQASEFPSELMSNDSKALCG"
     primer_bind     complement(454..478)
                     /label=Gibson-rev
     terminator      597..644
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(649..665)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    824..1074
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1077..1832)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      1923..2142
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    2562..2583
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2598..2628
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2636..2652
                     /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     2660..2676
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2749..3594
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    3702..3853
                     /label=nnpp promoters
     CDS             3893..4708
                     /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(4818..4861)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     4879..4895
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4896..4927
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgaaggcc
       61 ttcagccccg tgagaagcgt gagaaagaac agcctgagcg accacagcct gggcatcagc
      121 agaagcaaga cccccgtgga cgaccccatg agcctgctgt acaacatgaa cgactgctac
      181 agcaagctga aggagctggt gcccagcatc ccccagaaca agaaggtgag caagatggag
      241 atcctgcagc acgtgatcga ctacatcctg gacctgcaga tcgccctgga cagccacccc
      301 accatcgtga gcctgcacca ccagagaccc ggccagaacc aggccagcag aacccccctg
      361 accaccctga acaccgacat cagcatcctg agcctgcagg ccagcgagtt ccccagcgag
      421 ctgatgagca acgacagcaa ggccctgtgc ggcggatccg ccaccaactt cagcctgcta
      481 gttataaggg tccgatgccg ctgcattagc agtttattca tagggataac agggtaatgt
      541 cagccaaagt aatgaatgtc gctgaggtgt caatcgtcgg agccgctgag caataactag
      601 cataacccct tggggcctct aaacgggtct tgaggggttt tttgcatggt catagctgtt
      661 tcctgagagc ttggcaggtg tcgcgtatgc atcgatcacg tgtccacgtt ctttaatagt
      721 ggactcttgt tccaaactgg aacaacactc ggatcgatcc ggcgcgcacc gtgggaaaaa
      781 ctccaggtag aggtacacac gcggatagcc aattcagagt aataaactgt gataatcaac
      841 cctcatcaat gatgacgaac taacccccga tatcaggtca catgacgaag ggaaagagaa
      901 ggaaatcaac tgtgacaaac tgccctcaaa tttggcttcc ttaaaaatta cagttcaaaa
      961 agtatgagaa aatccatgca ggctgaagga aacagcaaaa ctgtgacaaa ttaccctcag
     1021 taggtcagaa caaatgtgac gaaccaccct caaatctgtg acagataacc ctcagactat
     1081 cctgtcgtca tggaagtgat atcgcggaag gaaaatacga tatgagtcgt ctggcggcct
     1141 ttctttttct caatgtatga gaggcgcatt ggagttctgc tgttgatctc attaacacag
     1201 acttgcagga agcggcggcg gaagtcaggc atacgctggt aactttgagg cagctggtaa
     1261 cgctctatga tccagtcgat tttcagagac acgatgcctg agccatccgg cttacgatac
     1321 tgacacaggg attcgtataa acgcatggca tacggattgg tgatttcttt tgtttcacta
     1381 agccgaaact gcgtaaaccg gttctgtaac ccgataaaga agggaatgag atatgggttg
     1441 atatgtacac tgtaaagccc tctggatgga ctgtgcgcac gtttgataaa ccaaggaaaa
     1501 gattcatagc ctttttcatc accggcatcc tcttcagggc gataaaaaac cacttccttc
     1561 cccgcgaaac tcttcaatgc ctgccgtata tccttactgg cttccgcaga ggtcaatccg
     1621 aatatttcag catatttagc aacatggatc tcgcagatac cgtcatgttc ctgtagggtg
     1681 ccatcagatt ttctgatctg gtcaacgaac agatacagca tacgtttttg atcccgggag
     1741 agactatatg ccgcctcagt gaggtcgttt gactggacga ttcgcgggct atttttacgt
     1801 ttcttgtgat tgataaccgc tgtttccgcc atgacagatc catgtgaagt gtgacaagtt
     1861 tttagattgt cacactaaat aaaaaagagt caataagcag ggataacttt gtgaaaaaac
     1921 agcttcttct gagggcaatt tgtcacaggg ttaagggcaa tttgtcacag acaggactgt
     1981 catttgaggg tgatttgtca cactgaaagg gcaatttgtc acaacacctt ctgtagaacc
     2041 agcatggata aaggcctaca aggcgctcta aaaaagaaga tctaaaaact ataaaaaaaa
     2101 taattataaa aatatccccg tggataagtg gataacccca agggaagttt tttcaggcat
     2161 cgtgtgtaag cagaatatat aagtgctgtt ccctggtgct tcctcgctca ctcgaaattc
     2221 ccggggatag ctttatgctt gtaaaccgtt ttgtgaaaaa atttttaaaa taaaaaaggg
     2281 gacctctagg gtccccaatt aattagtaat ataatctatt aaaggtcatt caaaaggtca
     2341 tccaccggat caattcccct gctcgcgcag gctgggtgcc aagctctcgg gtaacatcaa
     2401 ggcccgatcc ttggagccct tgccctcccg cacgatgatc gtgccgtgat cgaaatccag
     2461 atccttgacc cgcagttgca aaccctcact gatccgattc attaatgcag ctggcacgac
     2521 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     2581 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     2641 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcggtacc
     2701 tgtcgcggca acgcgctaac agacgtagta agaaccacca gcattgtaat gctggctaaa
     2761 gtcactttcc tgagctgtat aacgatgagc gattttactt tttctggcta tgaattggcc
     2821 tgctttgtaa cacactccgg tctatcccgt agcgccgggc atatcctgtc gcaatgtgca
     2881 aatctcgcgg caacaaccag tgaatacttc attcacaagc ctcaccgcct gatcgcggca
     2941 gaaactggtt atagccaatc aaccgtcgtt cgtgcattcc gtgaagctgt aaacaaaggt
     3001 atcctgtctg tagagattgt tatcggcgat caccgtgaac gtcgcgctaa cctgtaccgg
     3061 tttacaccat cctttttggc cttcgcacaa caagccaaaa atgcgctgat agaaagcaaa
     3121 ttaaagatct cttcagcggc aaccaaggtt aaagctgttc tcgctaagac attggcttta
     3181 tttaattttt tatccacacc cccatgtcaa aatgataccc cctccccctg tcaggatgac
     3241 gtggcaataa agaataagaa gtcacaagtt aaaaaaacaa aaagatcagt ttccggcggt
     3301 gccggaacaa ccagcctcaa aaaattgact tcatggatcg ctaaggcaaa agcaaaggct
     3361 gacaatctgc ggttatccaa aaaacgcact caaaaacatg agttcaagca gaaagtagag
     3421 gcggctgcgc ggaaatatgc ttacctgaag aacaagcgtt cgcctgatat tggcgggata
     3481 tcaaacttcg ataacctacc gcattgcatg acggtaaacg aagctcttaa tgcggtttta
     3541 gccaaaaata aagataacga acaatggggt ataccggcag gattcagagg gtaatgaatt
     3601 gctctaatta taaccatgca tactttcaac acctctagtt tgccatgagg caaactcata
     3661 ggtgtcctgg taagaggaca ctgttgccaa aactggacgc cccattattg caattaataa
     3721 acaactaacg gacaattcta cctaacaata agtggagttg cggccggccg cgatccgcta
     3781 gcaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3841 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3901 tattcaacgg gaaacgtctt gctcgcgccc gcgattaaat tccaacatgg atgctgattt
     3961 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     4021 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     4081 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     4141 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     4201 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     4261 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     4321 cgatcgcgta tttcgcctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     4381 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     4441 taaactgttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     4501 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     4561 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     4621 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata aattgcagtt
     4681 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggcg
     4741 agctctgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     4801 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
     4861 ctgcacctgc accagtcagt aaaacgacgg ccagtagtca aaagcctccg accggaggct
     4921 tttgacttgg ttcaggtgga gtgggag
//
