LOCUS       sequence-483856-        6492 bp    DNA     circular SYN 19-AUG-2025
DEFINITION  Encodes AT1R, a key receptor on RAAS that regulated blood pressure .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6492)
  TITLE     Zaid Abassi Lab Plasmids 2025
REFERENCE   2  (bases 1 to 6492)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 19, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: AT1R-HA
FEATURES             Location/Qualifiers
     source          1..6492
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(29..47)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(30..47)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(67..84)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     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"
     primer_bind     complement(430..449)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     640..661
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(780..800)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        785..1114
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      965..1100
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1027..1046
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1181..1975
                     /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(1235..1254)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1845..1864
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2149..2270
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2186..2205)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2240..2259
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2319..2335)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2319..2335)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2332..2354)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2343..2359
                     /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(join(2367..2373,2374..2391,2392..2397))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2412..2433
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2550..2567)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2721..3309)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2801..2820)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3480..4271,4272..4340))
                     /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     4103..4122
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4341..4445)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4520..4539)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4711..5090
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5091..5294
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5245..5265
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5339..5358
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5339..5357
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5380..5406
                     /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 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      541 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      601 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      661 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      721 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
      781 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      841 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
      901 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
      961 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     1021 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     1081 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     1141 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     1201 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     1261 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     1321 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     1381 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     1441 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     1501 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     1561 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     1621 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     1681 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     1741 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     1801 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     1861 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     1921 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     1981 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     2041 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     2101 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     2161 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2221 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     2281 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     2341 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2401 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2461 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2521 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2581 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2641 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2701 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2761 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2821 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2881 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2941 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3001 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3061 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3121 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3181 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3241 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3301 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3361 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3421 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3481 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3541 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3601 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3661 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3721 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3781 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3841 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3901 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3961 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4021 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4081 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4141 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4201 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4261 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4321 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4381 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4441 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4501 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4561 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4621 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4681 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4741 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4801 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4861 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4921 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4981 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     5041 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     5101 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     5161 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     5221 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     5281 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
     5341 atacgactca ctatagggag acccaagctt ggtaccatgt acccatacga tgttccagat
     5401 tacgctatga tcctcaactc ctcaactgaa gacggtataa aaagaatcca ggatgattgc
     5461 ccaaaagcag ggcggcataa ctacatattc gtgatgatcc ccactcttta tagtattatc
     5521 tttgtggtgg gcatcttcgg gaattcactc gttgtgatcg tgatttactt ttatatgaag
     5581 cttaagacgg tggcctccgt tttcctgctg aatctcgctt tggccgacct gtgctttctt
     5641 ctcaccttgc ccttgtgggc cgtgtacacc gcgatggagt accgatggcc tttcggtaac
     5701 tacctgtgta agatcgctag cgccagcgta tcttttaatc tttatgcctc tgttttcctc
     5761 ctcacatgcc tgtctattga tcggtatctg gccatcgtac atccgatgaa atcaagactt
     5821 aggaggacaa tgctggttgc taaggtaacc tgtattatta tttggctgct ggcaggtctg
     5881 gcttccctgc ctgcaattat tcatcgcaac gtctttttca tcgagaacac caacattacc
     5941 gtctgcgcct ttcactacga gagtcaaaat tccacgctcc caataggact gggcctgaca
     6001 aaaaacattc tcggttttct ctttccattc ctcatcatct tgacctcata taccttgatc
     6061 tggaaggcac tgaaaaaggc ctatgagatc cagaaaaaca aacccagaaa cgacgatatt
     6121 tttaaaatca tcatggctat tgtgctgttt tttttcttca gttggatccc acaccagatc
     6181 tttaccttcc ttgacgtctt gattcaactg ggcatcatca gagattgccg cattgccgac
     6241 atcgtggaca cggccatgcc cattacgatt tgcatcgcct actttaataa ctgccttaat
     6301 cccctgttct acgggttcct cggcaagaaa ttcaagcggt actttcttca gctgctcaaa
     6361 tacatcccgc ccaaggcaaa aagccactca aatctgtcaa ccaagatgag tacactgagt
     6421 tacagaccat ccgacaacgt gagcagttct accaagaaac ccgcaccatg tttcgaggtc
     6481 gaatgagcgg cc
//
