LOCUS       sequence-11265-e        7563 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 7563)
  TITLE     Lawrence Cohen lab plasmids
REFERENCE   2  (bases 1 to 7563)
  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: Zahra74-75, 80-81
FEATURES             Location/Qualifiers
     source          1..7563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2988..3005)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2994..3218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3264..3692
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3351..3370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3561..3582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3701..3721)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3706..4035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3886..4021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3948..3967
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4084..5109
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             4084..5109
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    5239..5360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5276..5295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5330..5349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5409..5425)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5409..5425)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5422..5444)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5433..5449
                     /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(5457..5463,5464..5481,5482..5487))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5502..5523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5640..5657)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5811..6396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5891..5910)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6567..7358,7359..7427))
                     /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     7190..7209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7428..7532)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg ccacgtctgt gcattttaac cctgggttag attccaaaga agttaatggt
      961 aattcggtga aagaagaggc agaggtgcag atcgatgatg gaaaggaaga aaccaaagac
     1021 ccagacacaa tgtaccatca ggtccggaaa aagataactc cttttgtgat gtcctttggt
     1081 tttcgtgttt ttggcctggt actgatcatc ttggacatca ttatggtcat tgtggacttg
     1141 tctctttctg agaagagtcg tgatgttgga ggtgcaccgg agactgtttc tctggtcatc
     1201 tctttctttt ttctcatcga tgtattgctc cgtgtttatg tggagggatt caaggtgtat
     1261 ttcagctcaa aactgaatat agtagatgct tgtattgtgg tcattactct ggtggtcacc
     1321 atgatctacg cattctccga tttctcagga gccagtctga ttccccaggt ggtgacattc
     1381 ctgaggtctc tgaggatcct aaggctggta cgcatcttca gactggcctc gcagaagaga
     1441 gagcttgaaa aagtcaccag gagaatggtt tcagagaaca aaagaagata tcgcatggtt
     1501 tctaagggtg aagagctgtt cactggggtg gtcccaattc tcgtggaact cgacggtgac
     1561 gtcaacggcc acaaattttc tgtctcagga gaaggagagg gtgacgcaac ctatgggaaa
     1621 ctcactctca aattcatctg cactaccgga aaactgccag tcccttggcc gacactggtc
     1681 actaccctca catggggggt gcaatgtttc gctcggtatc cagatcacat gaaacagcac
     1741 gacttcttca aaagtgcaat gcctgagggc tacgtccagg aacgcactat atttttcaag
     1801 gacgacggga attataaaac tagggcagag gtcaaatttg aaggagacac cctcgtcaac
     1861 cggattgaac tcaaaggtat tgactttaag gaggatggaa atatcctggg tcacaaactg
     1921 gaatacaatg ctatctcaga taacgtgtac atcacagccg acaagcaaaa gaatggcatc
     1981 aaggcaaatt tcaagattcg gcacaacatc gaagatggtt ccgtgcagct cgccgatcac
     2041 tatcagcaaa atactcccat tggtgatgga cccgtcctgc tccctgataa ccactacctg
     2101 tcaacacaat ctaaactcag taaggacccc aatgagaaac gtgaccacat ggtgctcctg
     2161 gaatttgtca ccgccgctgg gatcactctg gggtcaatgg tatcaaaggg tgaagaactc
     2221 ttcactggtg tcgtcccaat cctggttgaa ctcgacggcg acgtcaacgg gcacaaattt
     2281 agtgtctcag gggaagggga gggcgacgct acctacggaa aactgaccct caagttcatt
     2341 tgcaccactg ggaaactccc tgtgccctgg ccaactctgg tcactacatt tggatacggc
     2401 ctgatgtgct ttgccaggta tcctgaccac atgaagcagc acgatttctt taagtcagct
     2461 atgccggagg ggtacgtgca agagaggact atttttttca aggatgacgg aaattataaa
     2521 acaagagcag aagttaagtt tgaaggggac accctggtga acagaataga gctgaaaggc
     2581 atcgatttca aagaagatgg aaacattctc gggcataagc tggagtacaa ctacaacagc
     2641 cacaacgtct atatcatggc tgacaaacag aagaatggaa ttaaagtgaa cttcaaaatc
     2701 agacataata tcgaagatgg ctcagtgcag ctcgccgacc actaccaaca gaacactcca
     2761 attggtgatg ggcctgtcct gctcccagat aaccactacc tctcttatca gagcaagctg
     2821 tcaaaagatc cgaatgagaa aagagatcat atggttctcc tggaatttgt cacagcagct
     2881 ggaatcacac tcggcatgga cgagctctac aagtgatagt aaaactcgag ctgcagaggc
     2941 ctgcatgcaa gcttggtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     3001 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3061 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3121 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3181 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3241 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3301 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3361 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3421 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3481 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3541 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3601 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3661 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3721 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3781 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3841 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3901 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3961 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4021 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcagcac
     4081 gtgatgaaaa agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc
     4141 gacagcgtct ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc
     4201 gatgtaggag ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa
     4261 gatcgttatg tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac
     4321 attggggaat tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg
     4381 ttgcaagacc tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg
     4441 gatgcgatcg ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa
     4501 ggaatcggtc aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg
     4561 tatcactggc aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat
     4621 gagctgatgc tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc
     4681 ggctccaaca atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag
     4741 gcgatgttcg gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg
     4801 gcttgtatgg agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg
     4861 ccgcggctcc gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt
     4921 gacggcaatt tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc
     4981 ggagccggga ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat
     5041 ggctgtgtag aagtactcgc cgatagtgga aaccgacgcc ccagcactcg tccgagggca
     5101 aaggaatagc acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5161 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5221 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5281 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5341 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     5401 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     5461 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     5521 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     5581 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5641 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5701 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5761 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5821 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5881 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5941 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6001 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6061 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6121 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6181 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6241 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6301 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     6361 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6421 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6481 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6541 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6601 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6661 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6721 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6781 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6841 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6901 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6961 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7021 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7081 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7141 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7201 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7261 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7321 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7381 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7441 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7501 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7561 gtc
//
