LOCUS       Exported                6493 bp ds-DNA     circular SYN 15-FEB-2024
DEFINITION  pcDNA3.1Star-AGTR1-KpnI and NotI-HA tag-N-ter 6493bp ds-DNA circular
            SYN  20-APR-2022.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6493)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 2, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6493
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(29..47)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    73..297
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      343..771
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        785..1115
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      966..1101
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1182..1976
                     /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"
     polyA_signal    2150..2271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2320..2336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2344..2360
                     /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(2368..2398)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2413..2434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2722..3310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3481..4341)
                     /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"
     promoter        complement(4342..4446)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4712..5091
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5092..5295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        5340..5358
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5381..5407
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gctcgagcat gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct
       61 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
      121 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
      181 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
      241 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
      301 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
      361 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      421 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      481 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
      541 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      601 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      661 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
      721 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
      781 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
      841 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
      901 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
      961 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     1021 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     1081 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     1141 ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     1201 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     1261 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     1321 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     1381 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     1441 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     1501 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     1561 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     1621 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     1681 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     1741 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     1801 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     1861 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     1921 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     1981 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     2041 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     2101 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     2161 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     2221 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     2281 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2341 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2401 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2461 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     2521 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2581 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2641 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2701 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2761 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2821 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2881 cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag
     2941 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3001 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3061 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3121 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3181 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3241 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3301 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3361 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3421 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3481 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3541 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3601 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3661 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3721 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3781 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3841 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3901 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3961 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     4021 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4081 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     4141 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4201 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4261 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4321 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4381 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4441 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     4501 cccctatggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     4561 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     4621 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     4681 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     4741 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     4801 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     4861 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggactat
     4921 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     4981 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     5041 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     5101 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
     5161 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
     5221 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
     5281 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
     5341 aatacgactc actataggga gacccaagct tggtaccatg tacccatacg atgttccaga
     5401 ttacgctatg atcctcaact cctcaactga agacggtata aaaagaatcc aggatgattg
     5461 cccaaaagca gggcggcata actacatatt cgtgatgatc cccactcttt atagtattat
     5521 ctttgtggtg ggcatcttcg ggaattcact cgttgtgatc gtgatttact tttatatgaa
     5581 gcttaagacg gtggcctccg ttttcctgct gaatctcgct ttggccgacc tgtgctttct
     5641 tctcaccttg cccttgtggg ccgtgtacac cgcgatggag taccgatggc ctttcggtaa
     5701 ctacctgtgt aagatcgcta gcgccagcgt atcttttaat ctttatgcct ctgttttcct
     5761 cctcacatgc ctgtctattg atcggtatct ggccatcgta catccgatga aatcaagact
     5821 taggaggaca atgctggttg ctaaggtaac ctgtattatt atttggctgc tggcaggtct
     5881 ggcttccctg cctgcaatta ttcatcgcaa cgtctttttc atcgagaaca ccaacattac
     5941 cgtctgcgcc tttcactacg agagtcaaaa ttccacgctc ccaataggac tgggcctgac
     6001 aaaaaacatt ctcggttttc tctttccatt cctcatcatc ttgacctcat ataccttgat
     6061 ctggaaggca ctgaaaaagg cctatgagat ccagaaaaac aaacccagaa acgacgatat
     6121 ttttaaaatc atcatggcta ttgtgctgtt ttttttcttc agttggatcc cacaccagat
     6181 ctttaccttc cttgacgtct tgattcaact gggcatcatc agagattgcc gcattgccga
     6241 catcgtggac acggccatgc ccattacgat ttgcatcgcc tactttaata actgccttaa
     6301 tcccctgttc tacgggttcc tcggcaagaa attcaagcgg tactttcttc agctgctcaa
     6361 atacatcccg cccaaggcaa aaagccactc aaatctgtca accaagatga gtacactgag
     6421 ttacagacca tccgacaacg tgagcagttc taccaagaaa cccgcaccat gtttcgaggt
     6481 cgaatgagcg gcc
//
