LOCUS       Exported                6378 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000530 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6378)
  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..6378
                     /organism="BBF10K_000530 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1884
                     /codon_start=1
                     /product="Nuclear Receptor Subfamily 4 Group A Member 1"
                     /label=NR4A1
                     /translation="MWLAKACWSIQSEMPCIQAQYGTPAPSPGPRDHLASDPLTPEFIK
                     PTMDLASPEAAPAAPTALPSFSTFMDGYTGEFDTFLYQLPGTVQPCSSASSSASSTSSS
                     SATSPASASFKFEDFQVYGCYPGPLSGPVDEALSSSGSDYYGSPCSAPSPSTPSFQPPQ
                     LSPWDGSFGHFSPSQTYEGLRAWTEQLPKASGPPQPPAFFSFSPPTGPSPSLAQSPLKL
                     FPSQATHQLGEGESYSMPTAFPGLAPTSPHLEGSGILDTPVTSTKARSGAPGGSEGRCA
                     VCGDNASCQHYGVRTCEGCKGFFKRTVQKNAKYICLANKDCPVDKRRRNRCQFCRFQKC
                     LAVGMVKEVVRTDSLKGRRGRLPSKPKQPPDASPANLLTSLVRAHLDSGPSTAKLDYSK
                     FQELVLPHFGKEDAGDVQQFYDLLSGSLEVIRKWAEKIPGFAELSPADQDLLLESAFLE
                     LFILRLAYRSKPGEGKLIFCSGLVLHRLQCARGFGDWIDSILAFSRSLHSLLVDVPAFA
                     CLSALVLITDRHGLQEPRRVEELQNRIASCLKEHVAAVAGEPQPASCLSRLLGKLPELR
                     TLCTQGLQRIFYLKLEDLVPPPPIIDKIFMDTLPF"
     primer_bind     complement(1885..1909)
                     /label=Gibson-rev
     terminator      2028..2075
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2080..2096)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2255..2505
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(2508..3263)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      3354..3573
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3993..4014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4029..4059
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4067..4083
                     /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     4091..4107
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4180..5025
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    5133..5284
                     /label=nnpp promoters
     CDS             5324..6139
                     /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(6249..6292)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     6310..6326
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      6327..6358
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgtggctg
       61 gcaaaagcat gttggagtat tcaaagtgaa atgccgtgta tacaagcaca atatggtaca
      121 cctgctcctt ctcctggacc tcgggatcat ctcgcgtctg atccgctcac tccggaattt
      181 ataaaaccaa cgatggatct cgcatcacca gaagctgcgc cagcagcgcc aacggctctt
      241 ccgtcctttt caacatttat ggatggatat acaggggaat ttgatacttt cttgtatcaa
      301 ctcccaggaa ctgtccaacc gtgttctagt gcgtccagtt ccgcatcatc tacatcctca
      361 tcttctgcaa cgtcccctgc tagtgcttct tttaaatttg aagattttca agtctatgga
      421 tgttatccag gaccactttc tgggccggtc gatgaagcac tctctagttc tggttcagat
      481 tattatggat ctccatgttc agcgccaagt ccttctacgc cttcatttca accacctcaa
      541 ttgtcaccat gggatgggag ttttgggcat ttctctccat cacaaacata tgaaggattg
      601 cgggcttgga ctgaacaact tcctaaagcg tctgggccac ctcaaccccc agctttcttt
      661 agcttttccc caccaactgg gccatctcca agtttggctc aatccccact taaattgttt
      721 ccatctcaag ctactcatca attgggagaa ggagaatctt attccatgcc gacggctttt
      781 cctgggcttg ctccgacaag tccacatctc gaagggtccg ggattctcga tactcctgtc
      841 acaagtacaa aagcgcggtc aggggcaccg ggtggaagtg aagggcggtg tgcagtctgt
      901 ggagataatg catcttgtca acattatgga gtccgaactt gcgaaggttg taaagggttc
      961 tttaaaagga cagtccagaa gaatgcaaaa tatatttgtt tggcaaataa agattgtccg
     1021 gtcgataaac gtcgacggaa tcgatgtcaa ttttgtcgct ttcaaaagtg tctcgcggtc
     1081 gggatggtta aagaagtcgt taggacagat tctcttaaag gtcggcgcgg gcgccttcct
     1141 agtaaaccga aacaaccgcc ggatgcatca ccagcgaatc tgcttacttc cctcgtaagg
     1201 gctcatttgg attccgggcc ttctacggca aaactcgatt attccaaatt tcaagaactc
     1261 gtacttccac attttgggaa agaagatgca ggagatgtcc aacaatttta tgatttgctt
     1321 tcaggttccc tcgaagtcat acgcaaatgg gcagagaaaa ttccaggatt tgcagaattg
     1381 tctcctgcag atcaagatct cttgttggaa tctgcatttc ttgaactttt cattttgagg
     1441 ttggcatatc gttcaaaacc tggtgaaggt aaactcattt tctgttcagg gcttgtgctc
     1501 catcggttgc aatgtgcgag gggttttggt gattggattg attccattct cgcattttca
     1561 cggtctttgc attcacttct tgtcgatgtc cctgcatttg catgtctttc agcgcttgtc
     1621 ttgattacgg atcggcatgg attgcaagaa ccgaggcgag tagaagaact tcaaaatcga
     1681 attgcatctt gtcttaaaga acatgtagca gcagttgctg gagaaccaca acctgcatct
     1741 tgtctttcta ggcttcttgg taaacttcca gaactccgca ctctttgtac acaagggctt
     1801 caacgaattt tctatttgaa attggaagat ttggtcccgc ctcctccaat aattgataaa
     1861 atatttatgg acaccctgcc cttcggatcc gccaccaact tcagcctgct agtatctcag
     1921 caagggcaac ccctacatat cctgaaactc atagggataa cagggtaata ttgacaaata
     1981 cgaccggtgg cgctgaggtg tcaatcgtcg gagccgctga gcaataacta gcataacccc
     2041 ttggggcctc taaacgggtc ttgaggggtt ttttgcatgg tcatagctgt ttcctgagag
     2101 cttggcaggt gtcgcgtatg catcgatcac gtgtccacgt tctttaatag tggactcttg
     2161 ttccaaactg gaacaacact cggatcgatc cggcgcgcac cgtgggaaaa actccaggta
     2221 gaggtacaca cgcggatagc caattcagag taataaactg tgataatcaa ccctcatcaa
     2281 tgatgacgaa ctaacccccg atatcaggtc acatgacgaa gggaaagaga aggaaatcaa
     2341 ctgtgacaaa ctgccctcaa atttggcttc cttaaaaatt acagttcaaa aagtatgaga
     2401 aaatccatgc aggctgaagg aaacagcaaa actgtgacaa attaccctca gtaggtcaga
     2461 acaaatgtga cgaaccaccc tcaaatctgt gacagataac cctcagacta tcctgtcgtc
     2521 atggaagtga tatcgcggaa ggaaaatacg atatgagtcg tctggcggcc tttctttttc
     2581 tcaatgtatg agaggcgcat tggagttctg ctgttgatct cattaacaca gacttgcagg
     2641 aagcggcggc ggaagtcagg catacgctgg taactttgag gcagctggta acgctctatg
     2701 atccagtcga ttttcagaga cacgatgcct gagccatccg gcttacgata ctgacacagg
     2761 gattcgtata aacgcatggc atacggattg gtgatttctt ttgtttcact aagccgaaac
     2821 tgcgtaaacc ggttctgtaa cccgataaag aagggaatga gatatgggtt gatatgtaca
     2881 ctgtaaagcc ctctggatgg actgtgcgca cgtttgataa accaaggaaa agattcatag
     2941 cctttttcat caccggcatc ctcttcaggg cgataaaaaa ccacttcctt ccccgcgaaa
     3001 ctcttcaatg cctgccgtat atccttactg gcttccgcag aggtcaatcc gaatatttca
     3061 gcatatttag caacatggat ctcgcagata ccgtcatgtt cctgtagggt gccatcagat
     3121 tttctgatct ggtcaacgaa cagatacagc atacgttttt gatcccggga gagactatat
     3181 gccgcctcag tgaggtcgtt tgactggacg attcgcgggc tatttttacg tttcttgtga
     3241 ttgataaccg ctgtttccgc catgacagat ccatgtgaag tgtgacaagt ttttagattg
     3301 tcacactaaa taaaaaagag tcaataagca gggataactt tgtgaaaaaa cagcttcttc
     3361 tgagggcaat ttgtcacagg gttaagggca atttgtcaca gacaggactg tcatttgagg
     3421 gtgatttgtc acactgaaag ggcaatttgt cacaacacct tctgtagaac cagcatggat
     3481 aaaggcctac aaggcgctct aaaaaagaag atctaaaaac tataaaaaaa ataattataa
     3541 aaatatcccc gtggataagt ggataacccc aagggaagtt ttttcaggca tcgtgtgtaa
     3601 gcagaatata taagtgctgt tccctggtgc ttcctcgctc actcgaaatt cccggggata
     3661 gctttatgct tgtaaaccgt tttgtgaaaa aatttttaaa ataaaaaagg ggacctctag
     3721 ggtccccaat taattagtaa tataatctat taaaggtcat tcaaaaggtc atccaccgga
     3781 tcaattcccc tgctcgcgca ggctgggtgc caagctctcg ggtaacatca aggcccgatc
     3841 cttggagccc ttgccctccc gcacgatgat cgtgccgtga tcgaaatcca gatccttgac
     3901 ccgcagttgc aaaccctcac tgatccgatt cattaatgca gctggcacga caggtttccc
     3961 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     4021 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     4081 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcggtac ctgtcgcggc
     4141 aacgcgctaa cagacgtagt aagaaccacc agcattgtaa tgctggctaa agtcactttc
     4201 ctgagctgta taacgatgag cgattttact ttttctggct atgaattggc ctgctttgta
     4261 acacactccg gtctatcccg tagcgccggg catatcctgt cgcaatgtgc aaatctcgcg
     4321 gcaacaacca gtgaatactt cattcacaag cctcaccgcc tgatcgcggc agaaactggt
     4381 tatagccaat caaccgtcgt tcgtgcattc cgtgaagctg taaacaaagg tatcctgtct
     4441 gtagagattg ttatcggcga tcaccgtgaa cgtcgcgcta acctgtaccg gtttacacca
     4501 tcctttttgg ccttcgcaca acaagccaaa aatgcgctga tagaaagcaa attaaagatc
     4561 tcttcagcgg caaccaaggt taaagctgtt ctcgctaaga cattggcttt atttaatttt
     4621 ttatccacac ccccatgtca aaatgatacc ccctccccct gtcaggatga cgtggcaata
     4681 aagaataaga agtcacaagt taaaaaaaca aaaagatcag tttccggcgg tgccggaaca
     4741 accagcctca aaaaattgac ttcatggatc gctaaggcaa aagcaaaggc tgacaatctg
     4801 cggttatcca aaaaacgcac tcaaaaacat gagttcaagc agaaagtaga ggcggctgcg
     4861 cggaaatatg cttacctgaa gaacaagcgt tcgcctgata ttggcgggat atcaaacttc
     4921 gataacctac cgcattgcat gacggtaaac gaagctctta atgcggtttt agccaaaaat
     4981 aaagataacg aacaatgggg tataccggca ggattcagag ggtaatgaat tgctctaatt
     5041 ataaccatgc atactttcaa cacctctagt ttgccatgag gcaaactcat aggtgtcctg
     5101 gtaagaggac actgttgcca aaactggacg ccccattatt gcaattaata aacaactaac
     5161 ggacaattct acctaacaat aagtggagtt gcggccggcc gcgatccgct agcaaagcca
     5221 cgttgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat catcatgaac
     5281 aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg
     5341 ggaaacgtct tgctcgcgcc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
     5401 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa
     5461 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
     5521 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
     5581 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc
     5641 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
     5701 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
     5761 atttcgcctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
     5821 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactgtt
     5881 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
     5941 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
     6001 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     6061 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     6121 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc gagctctgcc
     6181 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6241 gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctgcacctg
     6301 caccagtcag taaaacgacg gccagtagtc aaaagcctcc gaccggaggc ttttgacttg
     6361 gttcaggtgg agtgggag
//
