LOCUS       sequence-350823-        7002 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 7002)
  AUTHORS   Luttrell LM, Ferguson SS, Daaka Y, Miller WE, Maudsley S, Della 
            Rocca GJ, Lin F, Kawakatsu H, Owada K, Luttrell DK, Caron MG, 
            Lefkowitz RJ
  TITLE     Beta-arrestin-dependent formation of beta2 adrenergic receptor-Src 
            protein kinase complexes.
  JOURNAL   Science. 1999 Jan 29. 283(5402):655-61.
  PUBMED    9924018
REFERENCE   2  (bases 1 to 7002)
  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: "Science. 
            1999 Jan 29. 283(5402):655-61."
COMMENT     Sequence Label: pcDNA3 c-SRC (WT)
FEATURES             Location/Qualifiers
     source          1..7002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(31..50)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     277..297
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(306..734)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(416..437)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     628..647
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    780..1004
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     993..1010
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        1030..1048
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1030..1047
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2721..2740)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2722..2740)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2785..2988)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2814..2834)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2989..3368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3540..3559
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3634..3738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3739..3807,3808..4599)
                     /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(3957..3976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4770..5358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5259..5278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5512..5529
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5646..5667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5682..5687,5688..5705,5706..5712)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5720..5736
                     /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     5725..5747
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5744..5760
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5744..5760
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5809..5930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5820..5839)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5874..5893
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(6104..6898)
                     /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(6215..6234)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6825..6844
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(6965..7002,1..292))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6979..7002,1..112)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta
       61 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
      121 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
      181 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
      241 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
      301 gaattaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
      361 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
      421 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
      481 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
      541 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
      601 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
      661 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
      721 gctacagggc gcgtggggat accccctaga gccccagctg gttctttccg cctcagaagc
      781 catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct cccccttgct
      841 gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat gcgatgcaat
      901 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
      961 gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg ctgatcagcg
     1021 agctctagca tttaggtgac actatagaat agggccctct agatgcatgc tcgagcggcc
     1081 gctctagcgc ctgtgcctag aggttctccc cgggctggta ctggggctcg gtggacgtga
     1141 agtagtcctc caggaaggcc tgcaggtact cgaaggtggg ccgctcctca ggctccttcc
     1201 gccagcactg gcacatgagg tcgtgcaggg actcgggaca ctccggcggg cagggcatcc
     1261 ggtagccccg ctccacctgg tccagcacct cgcggttcac catcccaggg tagggcaccc
     1321 gtccctttgt ggtgagctca gtcagcagga tcccgaagga ccacacgtcc gacttgatgg
     1381 tgaagcggcc atagagggca gcttctggag ccgtccactt gatggggaat ttggcacctt
     1441 gccgcgccgt gtactcattg tcttcaatga gccgagccag cccaaagtcg gccactttgc
     1501 acaccaggtt ctctcccacc aggatgttgg ctgcacgaag gtcccggtgg acgtagttca
     1561 tccgctccac gtacgccatg cctgaggcga tctgagcagc catgtccacc agctgaggca
     1621 gccgcaggta cttgcctgtc tcccccttga gaaagtccag caaactcccc ttgctcatgt
     1681 actccgtgac gatgtaaatg ggctcctctg aaaccacagc atacaactgc accagcttct
     1741 catgcctcag cttcttcatg acctgggcct cctgcaggaa ggcctctgga gacatcgtgc
     1801 caggcttcag ggttttgatg gccaccctgg tggtaccgtt ccaggtcccc atccacacct
     1861 cgccaaagca gccctggccc agcttgacct ccagccgcag cgactcccga gggatctccc
     1921 aggcatcctt ggccaggccc tgagtctgcg gcttggacgt ggggcacacg gtggtgaggc
     1981 ggtggcacag gccatcggcg tgtttggagt agtaggccac cagctgctgc aggctgttga
     2041 actgggtgcg ggaggtgatg tagaagccgc cgctgtccag cttgcggatc ttgtagtgct
     2101 tcacgttgag gcccttggcg ttgtcgaagt cagacactga gaggcagtag gcacctttcg
     2161 tggtctcact ttctcgcacg aagaaggtcc ctctcgggtt ctctgcattg agcagtaacc
     2221 gctctgactc ccgtctggtg atcttgccaa aataccactc ctcagcctgg atggagtcgg
     2281 agggcgccac gtagttgctg gggatgtagc ctgtctgtcc tgtgctgagc gagtgggcca
     2341 gccaccagtc tccctctgtg ttgttgacaa tctggagccg ctcgcctttc ttgaaggaca
     2401 ggtctgtctc cgtcctagac tcatagtcat agagggccac aaaggtggtc actccaccgg
     2461 ccagcgggcc cgccctctgc ggggaggtga cggtgtccga ggagttgaag cctccgaaca
     2521 gcttgggctc ggcggccgcg ggggcgaagg ccgcgctggg gccgcggtgg ccgtcggccg
     2581 aggctggctt gctgggggtc tgcgaggcgg ggaaagcgcc cccgccagcg ccgtgcacgt
     2641 tctcggcggg ctccaggctg cggcgccgct ggctggcatc cttgggcttg ctcttgttgc
     2701 tccccatgaa gcttgggtct ccctatagtg agtcgtatta atttcgataa gccagtaagc
     2761 agtgggttct ctagttagcc agagagctct gcttatatag acctcccacc gtacacgcct
     2821 accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt cccgttgatt
     2881 ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa atccccgtga
     2941 gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca tggtaatagc
     3001 gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca tgtactgggc
     3061 ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac ttggcatatg
     3121 atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc attgacgtca
     3181 atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg tcaatgggcg
     3241 ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa cgcggaactc
     3301 catatatggg ctatgaacta atgaccccgt aattgattac tattaataac tagtcaataa
     3361 tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct
     3421 aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt aaattttgct
     3481 cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc ttaactatgc
     3541 ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc tcccgatccg
     3601 tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3661 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3721 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3781 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3841 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3901 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3961 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4021 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4081 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4141 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4201 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4261 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     4321 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4381 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4441 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4501 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4561 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4621 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4681 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4741 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4801 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4861 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     4921 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4981 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5041 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5101 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5161 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5221 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5281 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5341 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5401 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5461 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5521 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     5581 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     5641 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     5701 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     5761 catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg
     5821 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     5881 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg
     5941 aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg
     6001 attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tgatggcagg
     6061 ttgggcgtcg cttggtcggt catttcgaac cccagagtcc cgctcagaag aactcgtcaa
     6121 gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa agcacgagga
     6181 agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc aacgctatgt
     6241 cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa aagcggccat
     6301 tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga tcctcgccgt
     6361 cgggcatgcg cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc tgatgctctt
     6421 cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct cgctcgatgc
     6481 gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc agccgccgca
     6541 ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac aggagatcct
     6601 gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca acgtcgagca
     6661 cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc tcgtcctgca
     6721 gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc ccctgcgctg
     6781 acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag tcatagccga
     6841 atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt tcaatcatgc
     6901 gaaacgatcc tcatcctgtc tcttgatcag atccgaaaat ggatatacaa gctcccggga
     6961 gctttttgca aaagcctagg cctccaaaaa agcctcctca ct
//
