LOCUS       Exported                5574 bp DNA     circular SYN 02-JUL-2021
DEFINITION  BBF10K_000508 in pOpen_v4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5574)
  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..5574
                     /organism="BBF10K_000508 in pOpen_v4"
     primer_bind     21..51
                     /label=Gibson-for
     CDS             52..1080
                     /codon_start=1
                     /product="Homo sapiens major histocompatibility complex, 
                     class I, G (HLA-G), transcript variant 1"
                     /label=HLAG
                     /translation="MKTPRMVVMAPRTLFLLLSGALTLTETWAGSHSMRYFSAAVSRPG
                     RGEPRFIAMGYVDDTQFVRFDSDSACPRMEPRAPWVEQEGPEYWEEETRNTKAHAQTDR
                     MNLQTLRGYYNQSEASSHTLQWMIGCDLGSDGRLLRGYEQYAYDGKDYLALNEDLRSWT
                     AADTAAQISKRKCEAANVAEQRRAYLEGTCVEWLHRYLENGKEMLQRADPPKTHVTHHP
                     VFDYEATLRCWALGFYPAEIILTWQRDGEDQTQDVELVETRPAGDGTFQKWAAVVVPSG
                     EEQRYTCHVQHEGLPEPLMLRWKQSSLPTIPIMGIVAGLVVLAAVVTGAAVAAVLWRKK
                     SSD"
     primer_bind     complement(1081..1105)
                     /label=Gibson-rev
     terminator      1224..1271
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1276..1292)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1451..1701
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             complement(1704..2459)
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     rep_origin      2550..2769
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     protein_bind    3189..3210
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3225..3255
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3263..3279
                     /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     3287..3303
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3376..4221
                     /codon_start=1
                     /label=repL
                     /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN
                     LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR
                     FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD
                     DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK
                     VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG
                     "
     misc_feature    4329..4480
                     /label=nnpp promoters
     CDS             4520..5335
                     /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(5445..5488)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     primer_bind     5506..5522
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5523..5554
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catgaagacc
       61 ccgcgcatgg tagtcatggc gccgaggaca cttttccttc tcttgtccgg agctctcact
      121 ctcacagaaa cttgggctgg tagtcattca atgcggtatt ttagcgcagc agttagtcga
      181 ccggggagag gagaacctcg ctttatagca atggggtatg tcgatgatac gcaatttgta
      241 cggtttgatt ccgattctgc ttgtccacga atggaaccgc gagctccgtg ggtcgaacaa
      301 gaagggcctg aatattggga agaagaaaca aggaatacta aagctcatgc tcaaacggat
      361 cgcatgaatt tgcaaacttt gaggggatat tacaatcaat cagaagcttc ttctcatact
      421 ttgcaatgga tgattggttg tgatcttgga tctgatggtc ggcttcttcg cgggtatgaa
      481 caatatgcat atgatgggaa agattatctt gcattgaatg aagatcttag gtcatggact
      541 gcagctgata cagctgctca aattagtaaa cgcaaatgtg aagcagcaaa tgtcgcggaa
      601 caacgccgcg catatttgga aggaacttgt gtcgaatggt tgcatcgcta tttggaaaat
      661 ggtaaagaaa tgctccaaag ggcagatcca ccgaaaacgc atgtcactca tcatcctgtc
      721 tttgattatg aagctacttt gcgctgttgg gctcttgggt tttatcctgc agaaattatt
      781 ttgacatggc aacgcgatgg agaagatcaa actcaagatg tcgaactcgt cgaaacgcgg
      841 ccggcaggtg atgggacatt tcaaaagtgg gcagctgttg tcgtcccttc tggtgaagaa
      901 caacggtata catgtcatgt tcaacatgaa gggctcccag aaccattgat gttgcgctgg
      961 aaacaatcct cacttccaac aattccaatt atgggaattg ttgcggggtt ggtcgtactc
     1021 gcggcagtag taacaggagc agcggttgca gcggtcctct ggagaaagaa gagcagcgac
     1081 ggatccgcca ccaacttcag cctgctagaa catagcattc acggggcctg ccacaattag
     1141 gacgggttag ggataacagg gtaataagtg gcctaccaac gagcgacgct gaggtgtcaa
     1201 tcgtcggagc cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     1261 ggggtttttt gcatggtcat agctgtttcc tgagagcttg gcaggtgtcg cgtatgcatc
     1321 gatcacgtgt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcgga
     1381 tcgatccggc gcgcaccgtg ggaaaaactc caggtagagg tacacacgcg gatagccaat
     1441 tcagagtaat aaactgtgat aatcaaccct catcaatgat gacgaactaa cccccgatat
     1501 caggtcacat gacgaaggga aagagaagga aatcaactgt gacaaactgc cctcaaattt
     1561 ggcttcctta aaaattacag ttcaaaaagt atgagaaaat ccatgcaggc tgaaggaaac
     1621 agcaaaactg tgacaaatta ccctcagtag gtcagaacaa atgtgacgaa ccaccctcaa
     1681 atctgtgaca gataaccctc agactatcct gtcgtcatgg aagtgatatc gcggaaggaa
     1741 aatacgatat gagtcgtctg gcggcctttc tttttctcaa tgtatgagag gcgcattgga
     1801 gttctgctgt tgatctcatt aacacagact tgcaggaagc ggcggcggaa gtcaggcata
     1861 cgctggtaac tttgaggcag ctggtaacgc tctatgatcc agtcgatttt cagagacacg
     1921 atgcctgagc catccggctt acgatactga cacagggatt cgtataaacg catggcatac
     1981 ggattggtga tttcttttgt ttcactaagc cgaaactgcg taaaccggtt ctgtaacccg
     2041 ataaagaagg gaatgagata tgggttgata tgtacactgt aaagccctct ggatggactg
     2101 tgcgcacgtt tgataaacca aggaaaagat tcatagcctt tttcatcacc ggcatcctct
     2161 tcagggcgat aaaaaaccac ttccttcccc gcgaaactct tcaatgcctg ccgtatatcc
     2221 ttactggctt ccgcagaggt caatccgaat atttcagcat atttagcaac atggatctcg
     2281 cagataccgt catgttcctg tagggtgcca tcagattttc tgatctggtc aacgaacaga
     2341 tacagcatac gtttttgatc ccgggagaga ctatatgccg cctcagtgag gtcgtttgac
     2401 tggacgattc gcgggctatt tttacgtttc ttgtgattga taaccgctgt ttccgccatg
     2461 acagatccat gtgaagtgtg acaagttttt agattgtcac actaaataaa aaagagtcaa
     2521 taagcaggga taactttgtg aaaaaacagc ttcttctgag ggcaatttgt cacagggtta
     2581 agggcaattt gtcacagaca ggactgtcat ttgagggtga tttgtcacac tgaaagggca
     2641 atttgtcaca acaccttctg tagaaccagc atggataaag gcctacaagg cgctctaaaa
     2701 aagaagatct aaaaactata aaaaaaataa ttataaaaat atccccgtgg ataagtggat
     2761 aaccccaagg gaagtttttt caggcatcgt gtgtaagcag aatatataag tgctgttccc
     2821 tggtgcttcc tcgctcactc gaaattcccg gggatagctt tatgcttgta aaccgttttg
     2881 tgaaaaaatt tttaaaataa aaaaggggac ctctagggtc cccaattaat tagtaatata
     2941 atctattaaa ggtcattcaa aaggtcatcc accggatcaa ttcccctgct cgcgcaggct
     3001 gggtgccaag ctctcgggta acatcaaggc ccgatccttg gagcccttgc cctcccgcac
     3061 gatgatcgtg ccgtgatcga aatccagatc cttgacccgc agttgcaaac cctcactgat
     3121 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     3181 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     3241 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     3301 gaccatgatt acgccaagcg cggtacctgt cgcggcaacg cgctaacaga cgtagtaaga
     3361 accaccagca ttgtaatgct ggctaaagtc actttcctga gctgtataac gatgagcgat
     3421 tttacttttt ctggctatga attggcctgc tttgtaacac actccggtct atcccgtagc
     3481 gccgggcata tcctgtcgca atgtgcaaat ctcgcggcaa caaccagtga atacttcatt
     3541 cacaagcctc accgcctgat cgcggcagaa actggttata gccaatcaac cgtcgttcgt
     3601 gcattccgtg aagctgtaaa caaaggtatc ctgtctgtag agattgttat cggcgatcac
     3661 cgtgaacgtc gcgctaacct gtaccggttt acaccatcct ttttggcctt cgcacaacaa
     3721 gccaaaaatg cgctgataga aagcaaatta aagatctctt cagcggcaac caaggttaaa
     3781 gctgttctcg ctaagacatt ggctttattt aattttttat ccacaccccc atgtcaaaat
     3841 gataccccct ccccctgtca ggatgacgtg gcaataaaga ataagaagtc acaagttaaa
     3901 aaaacaaaaa gatcagtttc cggcggtgcc ggaacaacca gcctcaaaaa attgacttca
     3961 tggatcgcta aggcaaaagc aaaggctgac aatctgcggt tatccaaaaa acgcactcaa
     4021 aaacatgagt tcaagcagaa agtagaggcg gctgcgcgga aatatgctta cctgaagaac
     4081 aagcgttcgc ctgatattgg cgggatatca aacttcgata acctaccgca ttgcatgacg
     4141 gtaaacgaag ctcttaatgc ggttttagcc aaaaataaag ataacgaaca atggggtata
     4201 ccggcaggat tcagagggta atgaattgct ctaattataa ccatgcatac tttcaacacc
     4261 tctagtttgc catgaggcaa actcataggt gtcctggtaa gaggacactg ttgccaaaac
     4321 tggacgcccc attattgcaa ttaataaaca actaacggac aattctacct aacaataagt
     4381 ggagttgcgg ccggccgcga tccgctagca aagccacgtt gtgtctcaaa atctctgatg
     4441 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     4501 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcttgct cgcgcccgcg
     4561 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
     4621 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
     4681 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
     4741 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
     4801 atggttactc accactgcga tccccgggaa aacagcattc caggtattag aagaatatcc
     4861 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
     4921 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgcctcgctc aggcgcaatc
     4981 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
     5041 tgttgaacaa gtctggaaag aaatgcataa actgttgcca ttctcaccgg attcagtcgt
     5101 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     5161 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     5221 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     5281 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcaga
     5341 attggttaat tggttgtaac actggcgagc tctgccacct gacgtctaag aaaccattat
     5401 tatcatgaca ttaacctata aaaataggcg tatcacgagg cagaatttca gataaaaaaa
     5461 atccttagct ttcgctaagg atgatttctg cacctgcacc agtcagtaaa acgacggcca
     5521 gtagtcaaaa gcctccgacc ggaggctttt gacttggttc aggtggagtg ggag
//
