LOCUS Exported 6145 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3 GFP LIC cloning vector (6D) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6145) TITLE 6-series mammalian LIC expression vectors REFERENCE 2 (bases 1 to 6145) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6145 /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" CDS 962..1678 /codon_start=1 /product="enhanced GFP (EGFP) derivative that exhibits robust folding and enhanced photostability (Zhong et al., 2019)" /label=SiriusGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(1004..1025) /label=EGFP-N /note="EGFP, reverse primer" primer_bind 1612..1633 /label=EGFP-C /note="EGFP, forward primer" promoter complement(1697..1715) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(1698..1715) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(1735..1752) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1741..1965 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2011..2439 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2098..2117) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2308..2329 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2448..2468) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2453..2783 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2634..2769 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2696..2715 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 2850..3644 /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(2904..2923) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3514..3533 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 3818..3939 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3855..3874) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3909..3928 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3988..4004) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3988..4004) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4001..4023) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4012..4028 /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(4036..4066) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4081..4102 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4219..4236) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4390..4978) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4470..4489) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5149..6009) /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 5772..5791 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6010..6114) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc 901 gagctcggat ccctacttcc aatccaatat tggtagtggg agcaacggca gcagcggatc 961 cgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 1021 cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg 1081 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct 1141 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca 1201 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatctcctt 1261 caaggacgac ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 1321 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa 1381 gctggagtac aacttcaaca gccacaacgt ctatatcacg gccgacaagc agaagaacgg 1441 catcaaggcg aacttcaaga tccgccacaa cgtcgaggac ggcagcgtgc agctcgccga 1501 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta 1561 cctgagcacc cagtccaagc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct 1621 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtagta 1681 atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga tcagcctcga 1741 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 1801 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 1861 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 1921 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa 1981 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg 2041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 2101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 2161 atcggggcat ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 2221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 2281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 2341 accctatctc ggtctattct tttgatttat aagggatttt ggggatttcg gcctattggt 2401 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca 2461 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc 2521 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2581 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 2641 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 2701 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 2761 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 2821 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 2881 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 2941 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3001 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3061 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3121 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3181 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 3241 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 3301 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 3361 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 3421 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 3481 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 3541 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 3601 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 3661 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 3721 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 3781 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 3841 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 3901 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 3961 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 4021 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 4081 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 4141 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 4201 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4261 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 4321 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 4381 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 4441 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 4501 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 4561 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt 4621 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 4681 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 4741 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 4801 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa 4861 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 4921 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 4981 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5041 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5101 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5161 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5221 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5281 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5341 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 5401 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 5461 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 5521 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 5581 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 5641 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 5701 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 5761 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 5821 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 5881 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 5941 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6001 aatactcata ctcttccttt ttcattatta ttgaagcatt tatcagggtt attgtctcat 6061 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6121 tccccgaaaa gtgccacctg acgtc //