LOCUS       sequence-372927-        7274 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7274)
  AUTHORS   Tang Y, Hu LA, Miller WE, Ringstad N, Hall RA, Pitcher JA, DeCamilli
            P, Lefkowitz RJ
  TITLE     Identification of the endophilins (SH3p4/p8/p13) as novel binding 
            partners for the beta1-adrenergic receptor.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Oct 26. 96(22):12559-64.
  PUBMED    10535961
REFERENCE   2  (bases 1 to 7274)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 1999 Oct 26. 96(22):12559-64."
COMMENT     Sequence Label: pcDNA3 Flag beta-2-adrenergic-receptor
FEATURES             Location/Qualifiers
     source          1..7274
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     168..185
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        205..223
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     205..222
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2168..2187)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2169..2187)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2232..2435)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2261..2281)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2436..2815
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     2987..3006
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3081..3185
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3186..3254,3255..4046)
                     /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(3404..3423)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4217..4805
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4706..4725
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4959..4976
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5093..5114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5129..5134,5135..5152,5153..5159)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5167..5183
                     /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     5172..5194
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5191..5207
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5191..5207
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5256..5377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5267..5286)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5321..5340
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5551..6345)
                     /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(5662..5681)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6272..6291
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6412..6741)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6426..6561
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6480..6499)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6726..6746
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(6755..7183)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6865..6886)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7077..7096
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    join(7229..7274,1..179)
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 atcctccccc ttgctgtcct gccccacccc accccccaga atagaatgac acctactcag
       61 acaatgcgat gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag
      121 ggtcaaggaa ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt
      181 cgaggctgat cagcgagctc tagcatttag gtgacactat agaatagggc cctctagatg
      241 catgctcgac tgctaccttt tttttttttt tcatggtcaa acaatttatt tactgtaaac
      301 tttagacttt gctcgggaaa acaactgaag ctgctcctca aatccctgcc taaaataccc
      361 cgtgtgagca aataagagga gtgggggaag agtccatgtt cacagctgaa aggtccgaga
      421 tactcaaaat cctcaagtcc aagggtgtag gaaggaaaat ctccttcaga tacaaattac
      481 cagcagcaat atattacccc taatacttga atagtgaggc agatacatgg aaaagtcatg
      541 aaaatatagc agactcaggt cctctaggac taaagctctt tagacataaa cttacaaatt
      601 ccatgcaaag aggaactgaa ctgtacaaat aacaaataat cactcaaata agttttctct
      661 ctttttacag cttaaaaaaa taaaaaaggc agaaggatgc ccttccttct gcatatctct
      721 atacaatttt acaattttat tctaagttta ttaccctcaa gttaaatagt ctgtttagtg
      781 ttctgttggg gggggggggn tctttaaaag tagaaaaact gctttacagc agtgagtcat
      841 ttgtactaca attcctccct tgtgaatcaa tgttatcgct aggcacagta ccttgatggc
      901 ccacaaagtc ttccgtgcct gggaggtctt cacacagcag tttattttct ttctcctgtt
      961 ccacgtgata tccactctgc tcccctgtgt tgccgttgct ggagtagcca ttcccatagg
     1021 ccttcaaaga agacctgcgc aggcacagaa gctcctggaa ggcaatcctg aaatctgggc
     1081 tccggcagta gataagggga ttgaaaccag aattgacata gcctatccaa tttaggagga
     1141 tgtaaacttc cttacggatg aggttatcct ggatcacatg cacaatgtta acgatgaaga
     1201 agggcagcca gcagagggtg aaagtgccca tgatgatgcc taacgtcttg agggctttgt
     1261 gctccttcaa gcagaacttg gaagatctgc ggagtccatg ccccgtccgc ccatcctgct
     1321 ccacctggct aaggttctgg acatggaagc ggccctcaga tttgtcaatc ttctggagct
     1381 gccttttggc ctcctgaaag accctggagt agacgaagac catgatcacc aggggaacgt
     1441 agaaggacac gatggaagag gcaatggcat aggcttggtt cgtgaagaag tcacagcagg
     1501 tctcattggc atagcagttg atggcttcct ggtgggtggc ccggtaccag tgcatctgaa
     1561 tgggcaagaa ggaggtaagg cctgacacaa tccacaccat cagaatgatc acccgggcct
     1621 tattcttggt cagcaggctc tggtacttga aaggtgaagt aatggcaaag tagcgatcca
     1681 ctgcgatcac gcacagggtc tcaatgctgg ccgtgacgca cagcacatca atggaagtcc
     1741 aaaactcgca ccagaagttg ccaaaagtcc acattttcat aagaatatgg gcggccccaa
     1801 agggcaccac tgccaggccc atgaccagat cagcacaggc cagtgaagtg atgaagtagt
     1861 tggtgaccgt ctgcagacgc tcgaacttgg caatggctgt gatgaccagc acattgccaa
     1921 acacgatggc caggacgatg agagacatga cgatgcccat gcccaccacc cacacctcgt
     1981 ccctctgctg cgtgacgtcg tggtccggcg catggcttct attgggtgcc agcaagaagg
     2041 cgctgccgtt cccgggttgc ccggcgtcat catcgtcctt gtagtcggcg aacaccaggc
     2101 agaagatgta gctcagggcg atgatggtct tcatggtgga tccgagctcg gtaccaagct
     2161 tgggtctccc tatagtgagt cgtattaatt tcgataagcc agtaagcagt gggttctcta
     2221 gttagccaga gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc
     2281 gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac
     2341 aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat
     2401 ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg
     2461 tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg
     2521 ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt
     2581 actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct
     2641 attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc
     2701 ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta
     2761 tgaactaatg accccgtaat tgattactat taataactag tcaataatca atgtcaacgc
     2821 gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag cagattcttc
     2881 atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc gcactactca
     2941 gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc atcagagcag
     3001 attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg acgtcaggtg
     3061 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3121 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3181 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3241 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3301 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3361 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3421 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3481 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3541 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3601 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3661 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3721 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3781 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3841 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3901 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3961 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4021 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4081 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4141 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4201 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4261 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actatttttc
     4321 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4381 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4441 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     4501 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     4561 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4621 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4681 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4741 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4801 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4861 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4921 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4981 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     5041 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     5101 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     5161 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     5221 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     5281 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     5341 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     5401 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     5461 acctttcata gaaggcggcg gtggaatcga aatctcgtga tggcaggttg ggcgtcgctt
     5521 ggtcggtcat ttcgaacccc agagtcccgc tcagaagaac tcgtcaagaa ggcgatagaa
     5581 ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc acgaggaagc ggtcagccca
     5641 ttcgccgcca agctcttcag caatatcacg ggtagccaac gctatgtcct gatagcggtc
     5701 cgccacaccc agccggccac agtcgatgaa tccagaaaag cggccatttt ccaccatgat
     5761 attcggcaag caggcatcgc catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc
     5821 cttgagcctg gcgaacagtt cggctggcgc gagcccctga tgctcttcgt ccagatcatc
     5881 ctgatcgaca agaccggctt ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg
     5941 gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc cgccgcattg catcagccat
     6001 gatggatact ttctcggcag gagcaaggtg agatgacagg agatcctgcc ccggcacttc
     6061 gcccaatagc agccagtccc ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg
     6121 aacgcccgtc gtggccagcc acgatagccg cgctgcctcg tcctgcagtt cattcagggc
     6181 accggacagg tcggtcttga caaaaagaac cgggcgcccc tgcgctgaca gccggaacac
     6241 ggcggcatca gagcagccga ttgtctgttg tgcccagtca tagccgaata gcctctccac
     6301 ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca atcatgcgaa acgatcctca
     6361 tcctgtctct tgatcagatc cgaaaatgga tatacaagct cccgggagct ttttgcaaaa
     6421 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg cagaggcggc
     6481 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     6541 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     6601 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     6661 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     6721 tccacaccct aactgacaca cattccacag aattaattcg cgttaaattt ttgttaaatc
     6781 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     6841 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     6901 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     6961 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     7021 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     7081 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     7141 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtggggata ccccctagag
     7201 ccccagctgg ttctttccgc ctcagaagcc atagagccca ccgcatcccc agcatgcctg
     7261 ctattgtctt ccca
//
