LOCUS Exported 6477 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses zebrafish TMEM206. In pcDNA3.1(+) backbone. ACCESSION . VERSION . KEYWORDS Zebrafish PAC (fPAC) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6477) AUTHORS Yang J, Chen J, Del Carmen Vitery M, Osei-Owusu J, Chu J, Yu H, Sun S, Qiu Z TITLE PAC, an evolutionarily conserved membrane protein, is a proton-activated chloride channel. JOURNAL Science. 2019 Apr 26;364(6438):395-399. doi: 10.1126/science.aav9739. Epub 2019 Apr 25. PUBMED 31023925 REFERENCE 2 (bases 1 to 6477) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6477 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2068..2085) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2074..2298 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2344..2772 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2431..2450) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2641..2662 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2781..2801) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2786..3115 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2966..3101 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3028..3047 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3182..3976 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3236..3255) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3846..3865 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 4150..4271 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4187..4206) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4241..4260 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4320..4336) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4320..4336) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4333..4355) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4344..4360 /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)." promoter complement(4368..4398) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4413..4434 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4551..4568) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4722..5310) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4802..4821) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5481..6341) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 6104..6123 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6342..6446) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc 961 agatatcatg ctgagaagag acacatcgcg ctcttatcag gagttcaatg gagaagagcg 1021 tcctgagaca gtagagtcca ttcgggacaa tggtgtttct gaagtggagc aagatgatga 1081 agcctgcgat cctttaccag atgaggacaa tgtggatagc aatctaatgc ccattggctt 1141 caacaaagct tgcctgaaga atgttttcac agtaatcctt gtcctcattt acttggcgct 1201 cacggcggtg gctgtgttct tggcctacca aaccatttca gacttcatgg acaaactcaa 1261 ccacccagtg atgtccgttt catacaaaga ggttgaagag tttgctgcac caggaattgt 1321 tttatttcct ggaaaggcac atttattaag ctgcatgcat cactatcacg acaacattcc 1381 acctcttgtt gcattggaaa atcttgcgaa aagagagtgt atgaaagagg aagtcatcta 1441 ccatgggccg tattcaaacc agacagaaaa gcgagctttg gtgttcagag ggccgactga 1501 cgtgaggaac cgagagctta tattccttca gcttagccga aacgaaacag aagaggattt 1561 cagcgcaatc agctatatga tttttgctaa attcagtgac atgcttgaaa gctcagataa 1621 agctgcattt atgatggact gtgaaaggaa ctactcgatg tggaccttct ctggaggctt 1681 taggacgtgg gtcaaaatgt ctttagtcag gacttcaggc agaagaaatg aatcagtaga 1741 atttagacag gagtctagtg tggtgaagta cattgacaaa aggcctccac tggagcaaac 1801 caacgagctt ttcttcatcg tgttccaatg gcgggatcca tttatacagc aagttaaaga 1861 catagtgaca gcaaacccgt ggaacaccat tgccattttg tgtggcgtct tcatggccct 1921 gtttaaagca gcagactttg ccaaactcag cattaagtgg atgatccgaa tccgcaaaag 1981 acacaaaagg gcaaaaatga gagaaatgaa ccaaatcagc taagcggccg ctcgagtcta 2041 gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg 2101 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt 2161 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg 2221 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg 2281 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc 2341 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga 2401 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg 2461 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat 2521 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg 2581 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata 2641 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt 2701 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat 2761 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc 2821 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa 2881 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 2941 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc 3001 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc 3061 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct 3121 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg 3181 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 3241 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 3301 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 3361 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 3421 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 3481 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 3541 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 3601 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 3661 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga 3721 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 3781 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 3841 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 3901 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 3961 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac 4021 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 4081 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 4141 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 4201 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 4261 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg 4321 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 4381 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 4441 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 4501 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact 4561 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta 4621 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag 4681 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc 4741 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta 4801 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg 4861 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc 4921 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac 4981 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac 5041 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg 5101 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga 5161 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt 5221 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag 5281 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct 5341 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg 5401 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat 5461 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc 5521 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg 5581 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct 5641 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca 5701 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg 5761 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg 5821 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc 5881 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag 5941 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg 6001 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag 6061 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat 6121 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg 6181 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca 6241 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca 6301 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat 6361 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag 6421 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc //