LOCUS       sequence-154547-        7562 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 7562)
  TITLE     src plasmids
REFERENCE   2  (bases 1 to 7562)
  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"
COMMENT     Sequence Label: pCMV5 mouse src K295R Y527F
FEATURES             Location/Qualifiers
     source          1..7562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(11..28)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1981..2000)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2172..2551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2552..2755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2706..2726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2800..2819
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2800..2818
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4958..4976)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4959..4976)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4996..5013)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5002..5226
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5272..5785
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5359..5378)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5569..5590
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5808..5828)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5813..6142
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5993..6128
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6209..7003
                     /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(6263..6282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6873..6892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7177..7311
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7214..7233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7268..7287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7342..7358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7342..7358)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7355..7377)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7366..7382
                     /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(7390..7396,7397..7414,7415..7420))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7435..7456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     1981 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     2041 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     2101 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     2161 atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2221 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2281 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2341 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2401 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     2461 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     2521 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     2581 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     2641 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     2701 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg
     2761 gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc actataggga
     2821 gacccaagct tatcgatgcc acagtggtac gcgaggccac cagcagagtc agctcagctc
     2881 ctgcagcaca cagacaaaag cgtatttcac aattaccaat agctgtcttt cctgccttcc
     2941 tcgattctag accactgtgg ccaggcggta gctgggacgt gcagccgacc accatgggca
     3001 gcaacaagag caagcccaag gacgccagcc agcggcgccg cagcctggag ccctcggaaa
     3061 acgtgcacgg ggcagggggc gccttcccgg cctcacagac accgagcaag cccgcctccg
     3121 ccgacggcca ccgcgggccc agcgccgcct tcgtgccgcc cgcggccgag cccaagctct
     3181 tcggaggctt caactcctcg gacaccgtca cctccccgca gagggcgggg cctctggcag
     3241 gtggggtgac cacctttgtg gccctctatg actatgagtc acggacagag actgacctgt
     3301 ccttcaagaa aggggagcgg ctgcagattg tcaataacac ggagggagac tggtggctgg
     3361 cacactcgct gagcacggga cagaccggtt acatccccag caactatgtg gcgccctccg
     3421 actccatcca ggctgaggag tggtactttg gcaagatcac tagacgggaa tcagagcggc
     3481 tgctgctcaa cgccgagaac ccgagaggga ccttcctcgt gagggagagt gagaccacaa
     3541 aaggtgccta ctgcctctct gtatccgact tcgacaatgc caagggccta aatgtgaaac
     3601 actacaagat ccgcaagctg gacagcggcg gtttctacat cacctcccgc acccagttca
     3661 acagcctgca gcagctcgtg gcttactact ccaaacatgc tgatggcctg tgtcaccgcc
     3721 tcactaccgt atgtcccaca tccaagcctc agacccaggg attggccaag gatgcgtggg
     3781 agatcccccg ggagtccctg cggctggagg tcaagctggg ccagggttgc ttcggagagg
     3841 tgtggatggg gacctggaac ggcaccacga gggttgccat caggactctg aagccaggca
     3901 ccatgtcccc agaggccttc ctgcaggagg cccaagtcat gaagaaactg aggcacgaga
     3961 aactggtgca gctgtatgct gtggtgtcgg aagaacccat ttacattgtg acagagtaca
     4021 tgaacaaggg gagtctgctg gactttctca agggggaaac gggcaaatat ttgcggctac
     4081 cccagctggt ggacatgtct gctcagatcg cttcaggcat ggcctatgtg gagcggatga
     4141 actatgtgca ccgggacctt cgagccgcca atatcctagt aggggagaac ctggtgtgca
     4201 aagtggccga ctttgggttg gcccggctca tagaagacaa cgaatacaca gcccggcaag
     4261 gtgccaaatt ccccatcaag tggaccgccc ctgaagctgc tctgtacggc aggttcacca
     4321 tcaagtcgga tgtgtggtcc tttgggattc tgctgaccga gctcaccact aagggaagag
     4381 tgccctatcc tgggatggtg aaccgtgagg ttctggacca ggtggagcgg ggctaccgga
     4441 tgccttgtcc ccccgagtgc cccgagtccc tgcatgacct tatgtgccag tgctggcgga
     4501 aggagcccga ggagcggccc accttcgagt acctgcaggc cttcctggaa gactacttta
     4561 cgtccactga gccacagttc cagcccgggg agaacctata gggactgtgt gcgcagaact
     4621 gtcacctcta ggcttccggg tgttgcctcc gatgcacctg cctcccttca tcctctctct
     4681 ggggctgcgt tgcctcggca atgccccttt tggagcatgg aagggcttca gggatggggc
     4741 agcctggaag gacaggtgga ggctggtaca gtgactgtcc catgcctgcc atggccatga
     4801 ttacgaattc gagctcgccc ggggatcctc tagagtcgac ctgcagccaa gcttggtacc
     4861 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca
     4921 ctggcggccg ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta
     4981 gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     5041 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     5101 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     5161 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     5221 ctatggcttc tgaggcggaa agaaccagct ggggctcgag gggggatccc cacgcgccct
     5281 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5341 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5401 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5461 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5521 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5581 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5641 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5701 ttaacaaaat attaacgctt acaatttaaa tatttgctta tacaatcttc ctgtttttgg
     5761 ggcttttctg attatcaacc ggggtgggta ccgagctcga attctgtgga atgtgtgtca
     5821 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5881 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     5941 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     6001 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     6061 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     6121 tggaggccta ggcttttgca aaaagctccc gggagcttgg atatccattt tcggatctga
     6181 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     6241 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     6301 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     6361 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     6421 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     6481 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     6541 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     6601 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     6661 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     6721 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     6781 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     6841 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     6901 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     6961 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     7021 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     7081 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     7141 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     7201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7261 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgtcgacct
     7321 cgagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     7381 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     7441 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     7501 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     7561 ct
//
