LOCUS       sequence-227071-        7080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse Src.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  AUTHORS   Schmitt JM, Stork PJ
  TITLE     PKA phosphorylation of Src mediates cAMP's inhibition of cell growth
            via Rap1.
  JOURNAL   Mol Cell. 2002 Jan;9(1):85-94.
  PUBMED    11804588
REFERENCE   2  (bases 1 to 7080)
  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: "Mol Cell.";
            date: "2002-01"; volume: "9(1)"; pages: "85-94"
COMMENT     Sequence Label: Flag-Src
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        123..502
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        503..706
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     657..677
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     751..770
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        751..769
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(785..801)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(786..802)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             2436..2483
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     promoter        complement(2521..2539)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2522..2539)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2559..2576)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2565..2789
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2835..3263
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2922..2941)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3132..3153
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3272..3292)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3277..3606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3457..3592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3519..3538
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3673..4467
                     /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(3727..3746)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4337..4356
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4641..4762
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4678..4697)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4732..4751
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4811..4827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4811..4827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4824..4846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4835..4851
                     /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(4859..4865,4866..4883,4884..4889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4904..4925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5042..5059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5213..5801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5293..5312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5972..6763,6764..6832))
                     /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     6595..6614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6833..6937)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7012..7031)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
       61 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatatacgcg
      121 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
      181 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
      241 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
      301 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
      361 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
      421 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
      481 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata
      541 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
      601 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
      661 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctct ggctaactag
      721 agaacccact gcttactggc ttatcgaaat taatacgact cactataggg agacccaagc
      781 ttatcgatac cgtcgacctc gagcggccgc cactgtgctg gatcatgggc agcaacaaga
      841 gcaagcccaa ggacgccagc cagcggcgcc gcagcctgga gccctcggaa aacgtgcacg
      901 gggcaggggg cgccttcccg gcctcacaga caccgagcaa gcccgcctcc gccgacggcc
      961 accgcgggcc cagcgccgcc ttcgtgccgc ccgcggccga gcccaagctc ttcggaggct
     1021 tcaactcctc ggacaccgtc acctccccgc agagggcggg gcctctggca ggtggggtga
     1081 ccacctttgt ggccctctat gactatgagt cacggacaga gactgacctg tccttcaaga
     1141 aaggggagcg gctgcagatt gttaacaaca cggagggaga ctggtggctg gcacactcgc
     1201 tgagcacggg acagaccggt tacatcccca gcaactatgt ggcgccctcc gactccatcc
     1261 aggctgagga gtggtacttt ggcaagatca ctagacggga atcagagcgg ctgctgctca
     1321 acgccgagaa cccgagaggg accttcctcg tgagggagag tgagaccaca aaaggtgcct
     1381 actgcctctc tgtatccgac ttcgacaatg ccaagggcct aaatgtgaaa cactacaaga
     1441 tccgcaagct ggacagcggc ggtttctaca tcacctcccg cacccagttc aacagcctgc
     1501 agcagctcgt ggcttactac tccaaacatg ctgatggcct gtgtcaccgc ctcactaccg
     1561 tatgtcccac atccaagcct cagacccagg gattggccaa ggatgcgtgg gagatccccc
     1621 gggagtccct gcggctggag gtcaagctgg gccagggttg cttcggagag gtgtggatgg
     1681 ggacctggaa cggcaccacg agggttgcca tcaaaactct gaagccaggc accatgtccc
     1741 cagaggcctt cctgcaggag gcccaagtca tgaagaaact gaggcacgag aaactggtgc
     1801 agctgtatgc tgtggtgtcg gaagaaccca tttacattgt gacagagtac atgaacaagg
     1861 ggagtctgct ggactttctc aagggggaaa cgggcaaata tttgcggcta ccccagctgg
     1921 tggacatgtc tgctcagatc gcttcaggca tggcctatgt ggagcggatg aactatgtgc
     1981 accgggacct tcgagccgcc aatatcctag taggggagaa cctggtgtgc aaagtggccg
     2041 actttgggtt ggcccggctc atagaagaca acgaatacac agcccggcaa ggtgccaaat
     2101 tccccatcaa gtggaccgcc cctgaagctg ctctgtacgg caggttcacc atcaagtcgg
     2161 atgtgtggtc ctttgggatt ctgctgaccg agctcaccac taagggaaga gtgccctatc
     2221 ctgggatggt gaaccgtgag gttctggacc aggtggagcg gggctaccgg atgccttgtc
     2281 cccccgagtg ccccgagtcc ctgcatgacc ttatgtgcca gtgctggcgg aaggagcccg
     2341 aggagcggcc caccttcgag tacctgcagg ccttcctgga agactacttt acgtccactg
     2401 agccacagta cgagcccggg gagaacctag gatccgacta caaggacgac gatgacaagg
     2461 actacaagga cgacgatgac aagtgagcgg ccgctcgagc atgcatctag agggccctat
     2521 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     2581 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2641 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2701 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2761 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     2821 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     2881 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     2941 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     3001 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     3061 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     3121 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     3181 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     3241 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3541 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3601 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     3661 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     3721 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg
     3781 ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc
     3841 ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct
     3901 tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa
     3961 gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg
     4021 gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa
     4081 gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat
     4141 gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg
     4201 cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc
     4261 atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac
     4321 cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg
     4381 gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc
     4441 tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag
     4501 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4561 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4621 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4681 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4741 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     5041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     5101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     5161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     5221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     5281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     5341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5641 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     6001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     6061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     6121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     6181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     6241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     6301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     6361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     6421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6961 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     7021 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
//
