LOCUS Exported 5368 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pAAVscCBPITuDlet-7Gpluc7x122BT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5368) TITLE Gao JOURNAL Unpublished REFERENCE 2 (bases 1 to 5368) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5368 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 912..1152 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 912..932 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 1083..1102 /label=LKO.1 5' /note="Human U6 promoter, forward primer" CDS 1439..1996 /codon_start=1 /product="secreted Gaussia luciferase" /label=hGLuc /note="human codon-optimized" /translation="MGVKVLFALICIAVAEAKPTENNEDFNIVAVASNFATTDLDADRG KLPGKKLPLEVLKEMEANARKAGCTRGCLICLSHIKCTPKMKKFIPGRCHTYEGDKESA QGGIGEAIVDIPEIPGFKDLEPMEQFIAQVDLCVDCTTGCLKGLANVQCSDLLKKWLPQ RCATFASKIQGQVDKIKGAGGD" primer_bind complement(2181..2198) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2187..2298 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 2418..2547 /label=AAV2 ITR primer_bind complement(2566..2583) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2566..2582) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2575..2597) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 2724..3179 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2811..2830) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3021..3042 /label=F1ori-F /note="F1 origin, forward primer" promoter 3205..3309 /gene="bla" /label=AmpR promoter CDS 3310..4170 /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 complement(3528..3547) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4341..4929 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4830..4849 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 5083..5100 /label=L4440 /note="L4440 vector, forward primer" protein_bind 5217..5238 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5253..5283 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5291..5307 /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 5296..5318 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 5315..5331 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 5315..5331 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" ORIGIN 1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtgtagcc atgctctagg 121 aagatcaatt cggtacaatt cacgcgtcga cattgattat tgactctggt cgttacataa 181 cttacggtaa atggcccgcc tggctgaccg cccaacgacc ccgcccattg acgtcaataa 241 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 301 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 361 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 421 gggactttcc tacttggcag tacatctact cgaggccacg ttctgcttca ctctccccat 481 ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc 541 gatgggggcg gggggggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg 601 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa 661 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc 721 gggcgggagc gggatcagcc accgcggtgg cggccctaga gtcgatcgag gaactgaaaa 781 accagaaagt taactggtaa gtttagtctt tttgtctttt atttcagccc cagtggaaag 841 acgcgcaggc aaaacgcacc acgtgacgga gcgtgaccgc gcgccgagcg cgcgccaagg 901 tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata cgatacaagg 961 ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta gtacaaaata 1021 cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta tgttttaaaa 1081 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttgggt ttatatatct 1141 tgtggaaagg acgcgggatc cgacggcgct aggatcatca acaactatac aaccatctta 1201 ctacctcaca agtattctgg tcacagaata caacaactat acaaccatct tactacctca 1261 caagatgatc ctagcgccgt cttttttaag ctgtcccgga tccggtggtg gtgcaaatca 1321 aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa gtgttacttc 1381 tgctctaaaa gctgcggaat tgtacccgcg gccgatccac cggtcgccac catctagcat 1441 gggagtcaaa gttctgtttg ccctgatctg catcgctgtg gccgaggcca agcccaccga 1501 gaacaacgaa gacttcaaca tcgtggccgt ggccagcaac ttcgcgacca cggatctcga 1561 tgctgaccgc gggaagttgc ccggcaagaa gctgccgctg gaggtgctca aagagatgga 1621 agccaatgcc cggaaagctg gctgcaccag gggctgtctg atctgcctgt cccacatcaa 1681 gtgcacgccc aagatgaaga agttcatccc aggacgctgc cacacctacg aaggcgacaa 1741 agagtccgca cagggcggca taggcgaggc gatcgtcgac attcctgaga ttcctgggtt 1801 caaggacttg gagcccatgg agcagttcat cgcacaggtc gatctgtgtg tggactgcac 1861 aactggctgc ctcaaagggc ttgccaacgt gcagtgttct gacctgctca agaagtggct 1921 gccgcaacgc tgtgcgacct ttgccagcaa gatccagggc caggtggaca agatcaaggg 1981 ggccggtggt gactagctcg acgctgatcg atccatcgcc gtgtaacaaa caccatacaa 2041 cactccacaa acaccataca acactccaca aacaccatac aacactccac aaacaccata 2101 caacactcca caaacaccat acaacactcc acaaacacca tacaacactc cacaaacacc 2161 atacaacact ccaggatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg 2221 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata 2281 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt 2341 ggggcaggac agcaaggggg aggattggga agacaattag gtagataagt agcatggcgg 2401 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc 2461 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc 2521 ggcctcagtg agcgagcgag cgcgcagcct taattaacct aattcactgg ccgtcgtttt 2581 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc 2641 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt 2701 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt 2761 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc 2821 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg 2881 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta 2941 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt 3001 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat 3061 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa 3121 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta 3181 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 3241 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 3301 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt 3361 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag 3421 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt 3481 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg 3541 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag 3601 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta 3661 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg 3721 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta 3781 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac 3841 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt 3901 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca 3961 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag 4021 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta 4081 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag 4141 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt 4201 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat 4261 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta 4321 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa 4381 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt 4441 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag 4501 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta 4561 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca 4621 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag 4681 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa 4741 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga 4801 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc 4861 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc 4921 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt 4981 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt 5041 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag 5101 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa 5161 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat 5221 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg 5281 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac 5341 gccagattta attaaggcct taattagg //