LOCUS Exported 8082 bp DNA circular SYN 02-JUL-2021 DEFINITION BBF10K_000541 in pOpen_v4. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8082) 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..8082 /organism="BBF10K_000541 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..3588 /codon_start=1 /product="ITGA1 Integrin subunit alpha 1" /label=CD49a /translation="MAPRPRARPGVAVACCWLLTVVLRCCVSFNVDVKNSMTFSGPVED MFGYTVQQYENEEGKWVLIGSPLVGQPKNRTGDVYKCPVGRGESLPCVKLDLPVNTSIP NVTEVKENMTFGSTLVTNPNGGFLACGPLYAYRCGHLHYTTGICSDVSPTFQVVNSIAP VQECSTQLDIVIVLDGSNSIYPWDSVTAFLNDLLERMDIGPKQTQVGIVQYGENVTHEF NLNKYSSTEEVLVAAKKIVQRGGRQTMTALGIDTARKEAFTEARGARRGVKKVMVIVTD GESHDNHRLKKVIQDCEDENIQRFSIAILGSYNRGNLSTEKFVEEIKSIASEPTEKHFF NVSDELALVTIVKTLGERIFALEATADQSAASFEMEMSQTGFSAHYSQDWVMLGAVGAY DWNGTVVMQKASQIIIPRNTTFNVESTKKNEPLASYLGYTVNSATASSGDVLYIAGQPR YNHTGQVIIYRMEDGNIKILQTLSGEQIGSYFGSILTTTDIDKDSNTDILLVGAPMYMG TEKEEQGKVYVYALNQTRFEYQMSLEPIKQTCCSSRQHNSCTTENKNEPCGARFGTAIA AVKDLNLDGFNDIVIGAPLEDDHGGAVYIYHGSGKTIRKEYAQRIPSGGDGKTLKFFGQ SIHGEMDLNGDGLTDVTIGGLGGAALFWSRDVAVVKVTMNFEPNKVNIQKKNCHMEGKE TVCINATVCFDVKLKSKEDTIYEADLQYRVTLDSLRQISRSFFSGTQERKVQRNITVRK SECTKHSFYMLDKHDFQDSVRITLDFNLTDPENGPVLDDSLPNSVHEYIPFAKDCGNKE KCISDLSLHVATTEKDLLIVRSQNDKFNVSLTVKNTKDSAYNTRTIVHYSPNLVFSGIE AIQKDSCESNHNITCKVGYPFLRRGEMVTFKILFQFNTSYLMENVTIYLSATSDSEEPP ETLSDNVVNISIPVKYEVGLQFYSSASEYHISIAANETVPEVINSTEDIGNEINIFYLI RKSGSFPMPELKLSISFPNMTSNGYPVLYPTGLSSSENANCRPHIFEDPFSINSGKKMT TSTDHLKRGTILDCNTCKFATITCNLTSSDISQVNVSLILWKPTFIKSYFSSLNLTIRG ELRSENASLVLSSSNQKRELAIQISKDGLPGRVPLWVILLSAFAGLLLLMLLILALWKI GFFKRPLKKKMEK" primer_bind complement(3589..3613) /label=Gibson-rev terminator 3732..3779 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind complement(3784..3800) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" misc_feature 3959..4209 /gene="incC" /label=incC /note="incompatibility region of the bacterial F plasmid" CDS complement(4212..4967) /codon_start=1 /gene="repE" /product="replication initiation protein for the bacterial F plasmid" /label=repE /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM RLSYIEKKKGRQTTHIVFSFRDITSMTTG" rep_origin 5058..5277 /label=ori2 /note="secondary origin of replication for the bacterial F plasmid; also known as oriS" protein_bind 5697..5718 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5733..5763 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5771..5787 /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 5795..5811 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" CDS 5884..6729 /codon_start=1 /label=repL /translation="MLAKVTFLSCITMSDFTFSGYELACFVTHSGLSRSAGHILSQCAN LAATTSEYFIHKPHRLIAAETGYSQSTVVRAFREAVNKGILSVEIVIGDHRERRANLYR FTPSFLAFAQQAKNALIESKLKISSAATKVKAVLAKTLALFNFLSTPPCQNDTPSPCQD DVAIKNKKSQVKKTKRSVSGGAGTTSLKKLTSWIAKAKAKADNLRLSKKRTQKHEFKQK VEAAARKYAYLKNKRSPDIGGISNFDNLPHCMTVNEALNAVLAKNKDNEQWGIPAGFRG " misc_feature 6837..6988 /label=nnpp promoters CDS 7028..7843 /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(7953..7996) /label=bacterial terminator /note="putative bacterial transcription terminator" primer_bind 8014..8030 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" terminator 8031..8062 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" ORIGIN 1 aagaattact gacccctcgg cagactgagt cgcccggggg gatccgccac catggcccct 61 cgtcctcgag ctaggccagg tgtagcagtc gcttgctgtt ggcttctgac tgtggtactt 121 agatgttgcg ttagcttcaa cgtggacgtg aagaacagca tgactttcag cggccccgtc 181 gaggacatgt tcgggtacac cgttcagcag tacgagaacg aggaaggtaa gtgggtcctg 241 atagggtcac ccttggtggg acagccgaag aaccgaaccg gtgacgttta caagtgccct 301 gtgggaaggg gagagtccct gccctgcgtt aagcttgacc tgccagttaa cacctcaatc 361 cccaacgtga cggaagtgaa ggagaacatg acattcggct ctacgctggt gactaacccc 421 aacggcggct tcttggcctg cggccctctc tacgcctacc ggtgcgggca cctgcactac 481 acaaccggca tttgctcaga cgtttccccc actttccagg tggtcaactc catcgctccc 541 gtgcaggagt gctctacgca gctggacatt gtgatcgtgt tggacggctc caactctatc 601 tacccttggg actccgtcac cgcttttctc aacgaccttc tggagaggat ggacatcggc 661 cccaagcaga ctcaggtggg tatcgtgcag tacggtgaga acgtcaccca cgagttcaac 721 ttgaacaagt acagctcaac agaagaggtt ctcgtggccg ccaagaagat cgtgcagcgg 781 ggcggacggc agacaatgac ggcacttggc attgacacag cacgaaagga agcctttaca 841 gaggccaggg gcgcccggcg cggggtgaag aaggtgatgg ttattgtgac tgacggagag 901 agtcacgaca accaccgtct gaagaaggtg atacaggatt gcgaagatga gaacatccag 961 agatttagta tcgccatatt gggatcttac aacaggggca accttagcac ggagaaattc 1021 gtcgaggaga tcaagtcaat cgcatctgag ccaacggaga aacacttctt caacgtgtct 1081 gacgagctgg cgttggttac aatcgtcaag accctgggtg agcgaatttt cgcccttgag 1141 gccaccgccg accagagtgc cgcaagcttc gagatggaga tgtcacagac tgggttcagc 1201 gcacactact cacaggactg ggtgatgctg ggagcagtgg gagcctacga ctggaacggt 1261 acagtggtga tgcaaaaggc aagccagatc atcatcccta gaaacactac attcaacgtg 1321 gagtctacca agaagaacga gcctctggct tcttacctgg ggtacaccgt gaactccgcc 1381 accgcatcta gcggcgacgt gctgtacatt gcgggccagc caaggtacaa ccacacaggg 1441 caggtgatca tctacagaat ggaagacgga aacataaaga tcctgcagac gctgagcggg 1501 gagcaaatcg gcagttactt cggaagcatc ctgacaacga ccgacattga caaggactct 1561 aacaccgaca tcctgctggt aggggccccc atgtacatgg gcacagagaa agaggagcag 1621 ggcaaggtgt acgtctacgc tttgaaccag acccgtttcg agtaccagat gagcctcgag 1681 ccaatcaagc agacatgctg ctcttcccga cagcacaact catgcactac cgagaacaag 1741 aacgagcctt gcggcgctcg cttcggtacc gccatcgcag cagttaagga cttgaacctg 1801 gacggcttca acgacatagt aatcggtgcc ccgcttgagg acgaccacgg cggcgcggtg 1861 tacatctacc acggatcagg aaagaccatc cgaaaggagt acgcgcagcg aatcccgagt 1921 ggcggggacg gcaagacact caagttcttc gggcagtcca tccacggtga gatggacctg 1981 aacggagacg ggcttacaga cgtgacaatc ggtggattgg gcggggcagc tcttttctgg 2041 tcaagggacg ttgctgtggt caaggtgacc atgaacttcg agcccaacaa ggtgaacatt 2101 cagaagaaga actgccacat ggaaggtaag gaaaccgtgt gcattaacgc taccgtatgc 2161 ttcgacgtga aactgaagag caaggaagac accatctacg aggccgacct gcagtaccgg 2221 gtgaccttgg acagcctgag gcagatttcc cgctctttct tcagcggaac ccaggagcgc 2281 aaggttcaga ggaacattac cgtgagaaag tcagagtgca ccaagcacag cttttacatg 2341 ctggacaagc acgacttcca ggacagcgtg agaattactc tggacttcaa cctgaccgac 2401 ccggagaacg gcccggtgct ggacgacagt ttgccaaact cagtgcacga gtacatacct 2461 ttcgccaagg actgcggcaa caaggaaaag tgcatatccg acctcagcct tcacgtggcg 2521 acaaccgaga aagacttgct tatcgtgcgt agccagaacg ataagttcaa cgtaagtctc 2581 actgttaaga acacgaagga ctccgcatac aacacacgaa ccatcgtgca ctacagtccc 2641 aaccttgttt tctctggcat agaggcaatc cagaaagatt cctgcgagag caaccacaac 2701 atcacctgca aggtgggcta ccccttcctg cgtcgaggtg agatggtgac cttcaagatc 2761 cttttccagt tcaatacgag ctacctgatg gagaacgtca ccatatacct cagcgctacc 2821 tcagactcag aggagccccc agagaccctt tccgacaacg tggtgaacat aagcatccca 2881 gtaaagtacg aggtgggact gcagttctac agttctgcta gcgagtacca catcagcatc 2941 gcagctaacg agaccgtgcc cgaggtgata aactctaccg aggacattgg caacgagatc 3001 aacatatttt acctgattcg aaagagtggc agtttcccca tgcccgagtt gaagctgagt 3061 attagtttcc ccaacatgac gtccaacggg taccccgtgc tctaccctac cgggctgagc 3121 agtagcgaga acgccaactg caggccacac atctttgagg accccttctc tataaactca 3181 ggaaagaaaa tgaccaccag caccgaccac cttaagcgcg gcacgattct tgactgcaac 3241 acctgcaagt tcgccacaat tacctgcaac ctcacgtcat cagacatctc ccaggtaaac 3301 gtctccttga tcctgtggaa gccaaccttc atcaagagct atttctcttc cctgaacctg 3361 acaatcaggg gcgagctgcg ctctgagaac gcctctctcg tgcttagttc ttccaaccaa 3421 aagcgagagc tggcaataca gattagcaag gacggcctgc ccggccgtgt ccccttgtgg 3481 gtgatcctgt tgtccgcttt cgccggcctg ctgcttctca tgctgctgat actggcactg 3541 tggaagatcg gcttcttcaa gcgtcctctg aagaagaaga tggagaaggg atccgccacc 3601 aacttcagcc tgctagttgt aatatcaccc gcgcactgca ccctcatggc ggtcctaggg 3661 ataacagggt aatttgatac tggaaatcga agaacgctga ggtgtcaatc gtcggagccg 3721 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc 3781 atggtcatag ctgtttcctg agagcttggc aggtgtcgcg tatgcatcga tcacgtgtcc 3841 acgttcttta atagtggact cttgttccaa actggaacaa cactcggatc gatccggcgc 3901 gcaccgtggg aaaaactcca ggtagaggta cacacgcgga tagccaattc agagtaataa 3961 actgtgataa tcaaccctca tcaatgatga cgaactaacc cccgatatca ggtcacatga 4021 cgaagggaaa gagaaggaaa tcaactgtga caaactgccc tcaaatttgg cttccttaaa 4081 aattacagtt caaaaagtat gagaaaatcc atgcaggctg aaggaaacag caaaactgtg 4141 acaaattacc ctcagtaggt cagaacaaat gtgacgaacc accctcaaat ctgtgacaga 4201 taaccctcag actatcctgt cgtcatggaa gtgatatcgc ggaaggaaaa tacgatatga 4261 gtcgtctggc ggcctttctt tttctcaatg tatgagaggc gcattggagt tctgctgttg 4321 atctcattaa cacagacttg caggaagcgg cggcggaagt caggcatacg ctggtaactt 4381 tgaggcagct ggtaacgctc tatgatccag tcgattttca gagacacgat gcctgagcca 4441 tccggcttac gatactgaca cagggattcg tataaacgca tggcatacgg attggtgatt 4501 tcttttgttt cactaagccg aaactgcgta aaccggttct gtaacccgat aaagaaggga 4561 atgagatatg ggttgatatg tacactgtaa agccctctgg atggactgtg cgcacgtttg 4621 ataaaccaag gaaaagattc atagcctttt tcatcaccgg catcctcttc agggcgataa 4681 aaaaccactt ccttccccgc gaaactcttc aatgcctgcc gtatatcctt actggcttcc 4741 gcagaggtca atccgaatat ttcagcatat ttagcaacat ggatctcgca gataccgtca 4801 tgttcctgta gggtgccatc agattttctg atctggtcaa cgaacagata cagcatacgt 4861 ttttgatccc gggagagact atatgccgcc tcagtgaggt cgtttgactg gacgattcgc 4921 gggctatttt tacgtttctt gtgattgata accgctgttt ccgccatgac agatccatgt 4981 gaagtgtgac aagtttttag attgtcacac taaataaaaa agagtcaata agcagggata 5041 actttgtgaa aaaacagctt cttctgaggg caatttgtca cagggttaag ggcaatttgt 5101 cacagacagg actgtcattt gagggtgatt tgtcacactg aaagggcaat ttgtcacaac 5161 accttctgta gaaccagcat ggataaaggc ctacaaggcg ctctaaaaaa gaagatctaa 5221 aaactataaa aaaaataatt ataaaaatat ccccgtggat aagtggataa ccccaaggga 5281 agttttttca ggcatcgtgt gtaagcagaa tatataagtg ctgttccctg gtgcttcctc 5341 gctcactcga aattcccggg gatagcttta tgcttgtaaa ccgttttgtg aaaaaatttt 5401 taaaataaaa aaggggacct ctagggtccc caattaatta gtaatataat ctattaaagg 5461 tcattcaaaa ggtcatccac cggatcaatt cccctgctcg cgcaggctgg gtgccaagct 5521 ctcgggtaac atcaaggccc gatccttgga gcccttgccc tcccgcacga tgatcgtgcc 5581 gtgatcgaaa tccagatcct tgacccgcag ttgcaaaccc tcactgatcc gattcattaa 5641 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat 5701 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg 5761 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac 5821 gccaagcgcg gtacctgtcg cggcaacgcg ctaacagacg tagtaagaac caccagcatt 5881 gtaatgctgg ctaaagtcac tttcctgagc tgtataacga tgagcgattt tactttttct 5941 ggctatgaat tggcctgctt tgtaacacac tccggtctat cccgtagcgc cgggcatatc 6001 ctgtcgcaat gtgcaaatct cgcggcaaca accagtgaat acttcattca caagcctcac 6061 cgcctgatcg cggcagaaac tggttatagc caatcaaccg tcgttcgtgc attccgtgaa 6121 gctgtaaaca aaggtatcct gtctgtagag attgttatcg gcgatcaccg tgaacgtcgc 6181 gctaacctgt accggtttac accatccttt ttggccttcg cacaacaagc caaaaatgcg 6241 ctgatagaaa gcaaattaaa gatctcttca gcggcaacca aggttaaagc tgttctcgct 6301 aagacattgg ctttatttaa ttttttatcc acacccccat gtcaaaatga taccccctcc 6361 ccctgtcagg atgacgtggc aataaagaat aagaagtcac aagttaaaaa aacaaaaaga 6421 tcagtttccg gcggtgccgg aacaaccagc ctcaaaaaat tgacttcatg gatcgctaag 6481 gcaaaagcaa aggctgacaa tctgcggtta tccaaaaaac gcactcaaaa acatgagttc 6541 aagcagaaag tagaggcggc tgcgcggaaa tatgcttacc tgaagaacaa gcgttcgcct 6601 gatattggcg ggatatcaaa cttcgataac ctaccgcatt gcatgacggt aaacgaagct 6661 cttaatgcgg ttttagccaa aaataaagat aacgaacaat ggggtatacc ggcaggattc 6721 agagggtaat gaattgctct aattataacc atgcatactt tcaacacctc tagtttgcca 6781 tgaggcaaac tcataggtgt cctggtaaga ggacactgtt gccaaaactg gacgccccat 6841 tattgcaatt aataaacaac taacggacaa ttctacctaa caataagtgg agttgcggcc 6901 ggccgcgatc cgctagcaaa gccacgttgt gtctcaaaat ctctgatgtt acattgcaca 6961 agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag 7021 gggtgttatg agccatattc aacgggaaac gtcttgctcg cgcccgcgat taaattccaa 7081 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc 7141 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa 7201 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt 7261 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac 7321 cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga 7381 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa 7441 ttgtcctttt aacagcgatc gcgtatttcg cctcgctcag gcgcaatcac gaatgaataa 7501 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt 7561 ctggaaagaa atgcataaac tgttgccatt ctcaccggat tcagtcgtca ctcatggtga 7621 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg 7681 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga 7741 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat 7801 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg 7861 gttgtaacac tggcgagctc tgccacctga cgtctaagaa accattatta tcatgacatt 7921 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt 7981 cgctaaggat gatttctgca cctgcaccag tcagtaaaac gacggccagt agtcaaaagc 8041 ctccgaccgg aggcttttga cttggttcag gtggagtggg ag //