LOCUS       Exported                5559 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000509 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5559)
  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..5559
                     /organism="BBF10K_000509 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     regulatory      46..55
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             52..1065
                     /codon_start=1
                     /product="Homo sapiens major histocompatibility complex, 
                     class I, G (HLA-G), transcript variant 1"
                     /label=HLAG1
                     /translation="MVVMAPRTLFLLLSGALTLTETWAGSHSMRYFSAAVSRPGRGEPR
                     FIAMGYVDDTQFVRFDSDSACPRMEPRAPWVEQEGPEYWEEETRNTKAHAQTDRMNLQT
                     LRGYYNQSEASSHTLQWMIGCDLGSDGRLLRGYEQYAYDGKDYLALNEDLRSWTAADTA
                     AQISKRKCEAANVAEQRRAYLEGTCVEWLHRYLENGKEMLQRADPPKTHVTHHPVFDYE
                     ATLRCWALGFYPAEIILTWQRDGEDQTQDVELVETRPAGDGTFQKWAAVVVPSGEEQRY
                     TCHVQHEGLPEPLMLRWKQSSLPTIPIMGIVAGLVVLAAVVTGAAVAAVLWRKKSSD"
     primer_bind     complement(1066..1090)
                     /label=Gibson-rev
     terminator      1209..1256
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1261..1277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1436..1686
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1689..2444)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2535..2754
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3174..3195
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3210..3240
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3248..3264
                     /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     3272..3288
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3361..4206
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4314..4465
                     /label=nnpp promoters
     CDS             4505..5320
                     /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(5430..5473)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5491..5507
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5508..5539
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catggtggtg
       61 atggcgccac ggacgttgtt tcttcttttg tccggagcgc ttactcttac agaaacgtgg
      121 gctggttcac atagtatgag gtatttctct gcggctgtct cacgccctgg acgtggtgaa
      181 cctcggttta tagctatggg atatgtcgat gatacgcaat ttgttcgctt tgattctgat
      241 agtgcttgtc ctcggatgga acctagggct ccttgggttg aacaagaagg tcctgaatat
      301 tgggaagaag aaactcggaa tactaaagcg catgcacaaa cagatcgaat gaatctccaa
      361 acgctccgtg gttattataa tcaaagtgaa gctagttccc atacacttca atggatgatt
      421 ggttgtgatc tcgggtccga tggacgcctt cttaggggat atgaacaata tgcatatgat
      481 ggaaaagatt atcttgcttt gaatgaagat cttcggtctt ggactgcagc agatacagcg
      541 gcgcaaattt caaaacggaa atgtgaagcg gcgaatgttg cagaacaaag gcgcgcgtat
      601 ctcgaaggaa catgtgtcga atggcttcat aggtatcttg aaaatgggaa agaaatgttg
      661 caacgcgccg atccccctaa aactcatgtt acacatcatc ctgtctttga ttatgaagct
      721 acgttgcgct gttgggctct tggtttctat cctgctgaaa ttattttgac atggcaacga
      781 gatggggaag atcaaactca agatgttgaa cttgtcgaaa caaggcctgc gggtgatggt
      841 acatttcaga aatgggcggc tgttgtagta ccgtccgggg aagaacaaag gtatacatgt
      901 catgtccaac atgaaggtct ccctgaacct ctcatgttgc ggtggaaaca atcctctctc
      961 ccaactattc caattatggg aattgtcgcg ggtctcgttg ttcttgcagc ggtcgttacg
     1021 ggagctgctg tcgcggctgt tctttggaga aagaagagca gcgacggatc cgccaccaac
     1081 ttcagcctgc tagacaacaa tctgaggctg gttcatggcc tacaaaacgc ggtagggata
     1141 acagggtaat tccagtgtga gctcaatttt tcgctgaggt gtcaatcgtc ggagccgctg
     1201 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgcatg
     1261 gtcatagctg tttcctgaga gcttggcagg tgtcgcgtat gcatcgatca cgtgtccacg
     1321 ttctttaata gtggactctt gttccaaact ggaacaacac tcggatcgat ccggcgcgca
     1381 ccgtgggaaa aactccaggt agaggtacac acgcggatag ccaattcaga gtaataaact
     1441 gtgataatca accctcatca atgatgacga actaaccccc gatatcaggt cacatgacga
     1501 agggaaagag aaggaaatca actgtgacaa actgccctca aatttggctt ccttaaaaat
     1561 tacagttcaa aaagtatgag aaaatccatg caggctgaag gaaacagcaa aactgtgaca
     1621 aattaccctc agtaggtcag aacaaatgtg acgaaccacc ctcaaatctg tgacagataa
     1681 ccctcagact atcctgtcgt catggaagtg atatcgcgga aggaaaatac gatatgagtc
     1741 gtctggcggc ctttcttttt ctcaatgtat gagaggcgca ttggagttct gctgttgatc
     1801 tcattaacac agacttgcag gaagcggcgg cggaagtcag gcatacgctg gtaactttga
     1861 ggcagctggt aacgctctat gatccagtcg attttcagag acacgatgcc tgagccatcc
     1921 ggcttacgat actgacacag ggattcgtat aaacgcatgg catacggatt ggtgatttct
     1981 tttgtttcac taagccgaaa ctgcgtaaac cggttctgta acccgataaa gaagggaatg
     2041 agatatgggt tgatatgtac actgtaaagc cctctggatg gactgtgcgc acgtttgata
     2101 aaccaaggaa aagattcata gcctttttca tcaccggcat cctcttcagg gcgataaaaa
     2161 accacttcct tccccgcgaa actcttcaat gcctgccgta tatccttact ggcttccgca
     2221 gaggtcaatc cgaatatttc agcatattta gcaacatgga tctcgcagat accgtcatgt
     2281 tcctgtaggg tgccatcaga ttttctgatc tggtcaacga acagatacag catacgtttt
     2341 tgatcccggg agagactata tgccgcctca gtgaggtcgt ttgactggac gattcgcggg
     2401 ctatttttac gtttcttgtg attgataacc gctgtttccg ccatgacaga tccatgtgaa
     2461 gtgtgacaag tttttagatt gtcacactaa ataaaaaaga gtcaataagc agggataact
     2521 ttgtgaaaaa acagcttctt ctgagggcaa tttgtcacag ggttaagggc aatttgtcac
     2581 agacaggact gtcatttgag ggtgatttgt cacactgaaa gggcaatttg tcacaacacc
     2641 ttctgtagaa ccagcatgga taaaggccta caaggcgctc taaaaaagaa gatctaaaaa
     2701 ctataaaaaa aataattata aaaatatccc cgtggataag tggataaccc caagggaagt
     2761 tttttcaggc atcgtgtgta agcagaatat ataagtgctg ttccctggtg cttcctcgct
     2821 cactcgaaat tcccggggat agctttatgc ttgtaaaccg ttttgtgaaa aaatttttaa
     2881 aataaaaaag gggacctcta gggtccccaa ttaattagta atataatcta ttaaaggtca
     2941 ttcaaaaggt catccaccgg atcaattccc ctgctcgcgc aggctgggtg ccaagctctc
     3001 gggtaacatc aaggcccgat ccttggagcc cttgccctcc cgcacgatga tcgtgccgtg
     3061 atcgaaatcc agatccttga cccgcagttg caaaccctca ctgatccgat tcattaatgc
     3121 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     3181 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     3241 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     3301 aagcgcggta cctgtcgcgg caacgcgcta acagacgtag taagaaccac cagcattgta
     3361 atgctggcta aagtcacttt cctgagctgt ataacgatga gcgattttac tttttctggc
     3421 tatgaattgg cctgctttgt aacacactcc ggtctatccc gtagcgccgg gcatatcctg
     3481 tcgcaatgtg caaatctcgc ggcaacaacc agtgaatact tcattcacaa gcctcaccgc
     3541 ctgatcgcgg cagaaactgg ttatagccaa tcaaccgtcg ttcgtgcatt ccgtgaagct
     3601 gtaaacaaag gtatcctgtc tgtagagatt gttatcggcg atcaccgtga acgtcgcgct
     3661 aacctgtacc ggtttacacc atcctttttg gccttcgcac aacaagccaa aaatgcgctg
     3721 atagaaagca aattaaagat ctcttcagcg gcaaccaagg ttaaagctgt tctcgctaag
     3781 acattggctt tatttaattt tttatccaca cccccatgtc aaaatgatac cccctccccc
     3841 tgtcaggatg acgtggcaat aaagaataag aagtcacaag ttaaaaaaac aaaaagatca
     3901 gtttccggcg gtgccggaac aaccagcctc aaaaaattga cttcatggat cgctaaggca
     3961 aaagcaaagg ctgacaatct gcggttatcc aaaaaacgca ctcaaaaaca tgagttcaag
     4021 cagaaagtag aggcggctgc gcggaaatat gcttacctga agaacaagcg ttcgcctgat
     4081 attggcggga tatcaaactt cgataaccta ccgcattgca tgacggtaaa cgaagctctt
     4141 aatgcggttt tagccaaaaa taaagataac gaacaatggg gtataccggc aggattcaga
     4201 gggtaatgaa ttgctctaat tataaccatg catactttca acacctctag tttgccatga
     4261 ggcaaactca taggtgtcct ggtaagagga cactgttgcc aaaactggac gccccattat
     4321 tgcaattaat aaacaactaa cggacaattc tacctaacaa taagtggagt tgcggccggc
     4381 cgcgatccgc tagcaaagcc acgttgtgtc tcaaaatctc tgatgttaca ttgcacaaga
     4441 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
     4501 tgttatgagc catattcaac gggaaacgtc ttgctcgcgc ccgcgattaa attccaacat
     4561 ggatgctgat ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac
     4621 aatctatcga ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg
     4681 tagcgttgcc aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat
     4741 gcctcttccg accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac
     4801 tgcgatcccc gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa
     4861 tattgttgat gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg
     4921 tccttttaac agcgatcgcg tatttcgcct cgctcaggcg caatcacgaa tgaataacgg
     4981 tttggttgat gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg
     5041 gaaagaaatg cataaactgt tgccattctc accggattca gtcgtcactc atggtgattt
     5101 ctcacttgat aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg
     5161 agtcggaatc gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt
     5221 ttctccttca ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa
     5281 taaattgcag tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt
     5341 gtaacactgg cgagctctgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5401 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
     5461 taaggatgat ttctgcacct gcaccagtca gtaaaacgac ggccagtagt caaaagcctc
     5521 cgaccggagg cttttgactt ggttcaggtg gagtgggag
//
