LOCUS       sequence-11434-e        8174 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 8174)
  TITLE     pN3-Sp1FL
REFERENCE   2  (bases 1 to 8174)
  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: pN3-Sp1FL
FEATURES             Location/Qualifiers
     source          1..8174
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    4962..5083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4999..5018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5053..5072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5090..5545)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5227..5248)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5439..5458
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5572..5676
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5678..6035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5678..5869
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5701..5721)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5886..6021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6070..6864
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             6070..6864
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6124..6143)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6734..6753
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7052..7071)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7096..7143
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7472..8060
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7961..7980
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgaggg ggccgccttg agcttgtccc tcagctgcca ccatgagcga
      661 ccaagatcac tccatggatg aaatgacagc tgtggtgaaa attgaaaaag gagttggtgg
      721 caataatggg ggcaatggta atggtggtgg tgccttttca caggctcgaa gtagcagcac
      781 aggcagtagc agcagcactg gaggaggagg gcaggagtcc cagccatccc ctttggctct
      841 gctggcagca acttgcagca gaattgagtc acccaatgag aacagcaaca actcccaggg
      901 cccgagtcag tcagggggaa caggtgagct tgacctcaca gccacacaac tttcacaggg
      961 tgccaatggc tggcagatca tctcttcctc ctctggggct acccctacct caaaggaaca
     1021 gagtggcagc agtaccaatg gcagcaatgg cagtgagtct tccaagaatc gcacagtctc
     1081 tggtgggcag tatgttgtgg ctgccgctcc caacttacag aaccagcaag ttctgacagg
     1141 actacctgga gtgatgccta atattcagta tcaagtaatc ccacagttcc agaccgttga
     1201 tgggcaacag ctgcagtttg ctgccactgg ggcccaagtg cagcaggatg gttctggtca
     1261 aatacagatc ataccaggtg caaaccaaca gattatcaca aatcgaggaa gtggaggcaa
     1321 catcattgct gctatgccaa acctactcca gcaggctgtc cccctccaag gcctggctaa
     1381 taatgtactc tcaggacaga ctcagtatgt gaccaatgta ccagtggccc tgaatgggaa
     1441 catcaccttg ctacctgtca acagcgtttc tgcagctacc ttgactccca gctctcaggc
     1501 agtcacgatc agcagctctg ggtcccagga gagtggctca cagcctgtca cctcagggac
     1561 taccatcagt tctgccagct tggtatcatc acaagccagt tccagctcct ttttcaccaa
     1621 tgccaatagc tactcaacta ctactaccac cagcaacatg ggaattatga actttactac
     1681 cagtggatca tcagggacca actctcaagg ccagacaccc cagagggtca gtgggctaca
     1741 ggggtctgat gctctgaaca tccagcaaaa ccagacatct ggaggctcat tgcaagcagg
     1801 ccagcaaaaa gaaggagagc aaaaccagca gacacagcag caacaaattc ttatccagcc
     1861 tcagctagtt caagggggac aggccctcca ggccctccaa gcagcaccat tgtcagggca
     1921 gacctttaca actcaagcca tctcccagga aaccctccag aacctccagc ttcaggctgt
     1981 tccaaactct ggtcccatca tcatccggac accaacagtg gggcccaatg gacaggtcag
     2041 ttggcagact ctacagctgc agaacctcca agttcagaac ccacaagccc aaacaatcac
     2101 cttagcccca atgcagggtg tttccttggg gcagaccagc agcagcaaca ccactctcac
     2161 acccattgcc tcagctgctt ccattcctgc tggcacagtc actgtgaatg ctgctcaact
     2221 ctcctccatg ccaggcctcc agaccattaa cctcagtgca ttgggtactt caggaatcca
     2281 ggtgcaccca attcaaggcc tgccgttggc tatagcaaat gccccaggtg atcatggagc
     2341 tcagcttggt ctccatgggg ctggtggtga tggaatacat gatgacacag caggtggaga
     2401 ggaaggagaa aacagcccag atgcccaacc ccaagccggt cggaggaccc ggcgggaagc
     2461 atgcacctgc ccctactgta aagacagtga aggaaggggc tcgggggatc ctggcaaaaa
     2521 gaaacagcat atttgccaca tccaaggctg tgggaaagtg tatggcaaga cctctcacct
     2581 gcgggcacac ttgcgctggc atacaggcga gaggccattt atgtgtacct ggtcatactg
     2641 tgggaaacgc ttcacacgtt cggatgagct acagaggcac aaacgtacac acacaggtga
     2701 gaagaaattt gcctgccctg agtgtcctaa gcgcttcatg aggagtgacc acctgtcaaa
     2761 acatatcaag acccaccaga ataagaaggg aggcccaggt gtagctctga gtgtgggcac
     2821 tttgcccctg gacagtgggg caggttcaga aggcagtggc actgccactc cttcagccct
     2881 tattaccacc aatatggtag ccatggaggc catctgtcca gagggcattg cccgtcttgc
     2941 caacagtggc atcaacgtca tgcaggtggc agatctgcag tccattaata tcagtggcaa
     3001 tggcttctga gatcaggcac ccggggccag agacatatgg gccatacccc ttaaccccgg
     3061 gatgcaaggt agcatgggtc caagagacat ggaagagaga gccatgaagc attaaaatgc
     3121 atggtgttga gaagaatcag gagagggata caagagagga gatggggtcc cggcacccat
     3181 ctgtatcatc agtgcctctt tgaaggtggg aaacattagt gaaaattctg ttggtgccac
     3241 gctttgatga gcatttgttt gaccccagtt tcttcttaca cttcttaccc cagcctaccc
     3301 ttcctgcatt tctcttctca gctcttccat gatggattcc cccccctttc ctaaagccat
     3361 catgccttga taaatatata tgatcattga aatacttttt aacaaaaaac agattctata
     3421 ttattatata tatatatata tataaagata tatagagatg cattcacagg gttggctgga
     3481 gaggaagacc attctgtgac caaaatacct tggtcatttt ttttatattg ccttatttcc
     3541 ctatggctga gccttgttgt gacacatcaa gcttttctgt agatgttgtc ttggcttccc
     3601 accagcttaa gcgttcatat gctctgcttt tagttcatat atacatacat aatgtttttc
     3661 ctttcttaat tttgtctttt tgtttgggat cagcttcttg cactccttcc ctaactcaac
     3721 tgttgccgtc tcatcttctc tcatctgatc acttcatgtt ttgtttttgt tactgcctgg
     3781 atgaggcact tctgtcaatt ttttcaggac cttagttcca gcagcagaat ggaaaaatcc
     3841 ttgaagccca ggctgatgct tgaagtaact gtggagggag tgttcaaaat actactgacg
     3901 caggcacctt cttggcgctg gagagtcaaa ggcatctccc ttcattagct gctctgagca
     3961 tcaagaatta gaagtctttc agtggaattg tacaagagtc cctttgaaga taataatctt
     4021 ggctcagttt gtataaactg tcaaattttc aaataatagg tagggggctt tcactaggaa
     4081 aatcatgtgc tcagaagagg aaatgactcg tagtcaggtt caggagttag tggagtattt
     4141 ggactttggt actgctgtct tccaaggtag ctctaagttt tgatgtgtgg gcttctgagt
     4201 ttatattctg aaaggaaata cacttctttt gaacatcccc actaggttct tttccattgt
     4261 caataaggag catcagccag tgaatctgtt tcaggtttcc attctgcaga actcctccaa
     4321 agcatgtgct agtggcaaga cagtggttct tatgatgttt tcccttaact tttccttgta
     4381 tgttcttggg tggttcctaa gggaaaggga agcacatgat catgggaatg atagcccaga
     4441 acaaaaagaa atcttgtctt accacagtgt tttataggag agattgggag aaatcatcct
     4501 gttttctctg tgacctgatt tcagaagaga ctgatccaaa aattataacg gcagggaacc
     4561 tagtgcattt ggcactgaga tttaaatgca accagaattg tcctcaaggc ccagccataa
     4621 aagcattgtc tctctcgacc ttctggtatc ttgttagaga gctttttact gtgaggaagt
     4681 gtggaaaaat aagctctgtg tgtgtgtgtg tgtgtgtgtg tgtgtaatct gttaggttgg
     4741 ggataggttt tctgctagcc aatattaaaa gagacctgca ccggaattcc ctcgagctca
     4801 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tcggccgcga ctctagatca
     4861 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4921 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4981 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     5101 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5161 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5221 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5281 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5341 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5401 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5461 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5521 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5581 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5641 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     5701 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5761 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5881 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     5941 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6001 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     6061 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6121 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6181 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6241 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6301 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6361 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6421 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6481 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6541 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     6601 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6661 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6721 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6781 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6841 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6901 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     6961 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7021 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     7081 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     7141 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     7201 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     7261 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     7321 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7381 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7441 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7501 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7561 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     7621 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7681 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7741 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7801 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7861 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7921 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     7981 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8041 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8101 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8161 ttaccgccat gcat
//
