LOCUS       sequence-209002-        8176 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 8176)
  TITLE     pN3-Sp1FL
REFERENCE   2  (bases 1 to 8176)
  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..8176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        119..422
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        423..626
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     577..597
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    5022..5143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5059..5078)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5113..5132
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5150..5605)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5287..5308)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5499..5518
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5632..5736
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5738..6095
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5738..5929
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5761..5781)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5946..6081
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6130..6924
                     /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             6130..6924
                     /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(6184..6203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6794..6813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7112..7131)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7156..7203
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7532..8120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8021..8040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta
       61 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      121 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      181 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      241 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      301 gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      361 tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      421 tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat
      481 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      541 actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac
      601 ggtgggaggt ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcgctacc
      661 ggactcagat ctcgaggggg ccgccttgag cttgtccctc agctgccacc atgagcgacc
      721 aagatcactc catggatgaa atgacagctg tggtgaaaat tgaaaaagga gttggtggca
      781 ataatggggg caatggtaat ggtggtggtg ccttttcaca ggctcgaagt agcagcacag
      841 gcagtagcag cagcactgga ggaggagggc aggagtccca gccatcccct ttggctctgc
      901 tggcagcaac ttgcagcaga attgagtcac ccaatgagaa cagcaacaac tcccagggcc
      961 cgagtcagtc agggggaaca ggtgagcttg acctcacagc cacacaactt tcacagggtg
     1021 ccaatggctg gcagatcatc tcttcctcct ctggggctac ccctacctca aaggaacaga
     1081 gtggcagcag taccaatggc agcaatggca gtgagtcttc caagaatcgc acagtctctg
     1141 gtgggcagta tgttgtggct gccgctccca acttacagaa ccagcaagtt ctgacaggac
     1201 tacctggagt gatgcctaat attcagtatc aagtaatccc acagttccag accgttgatg
     1261 ggcaacagct gcagtttgct gccactgggg cccaagtgca gcaggatggt tctggtcaaa
     1321 tacagatcat accaggtgca aaccaacaga ttatcacaaa tcgaggaagt ggaggcaaca
     1381 tcattgctgc tatgccaaac ctactccagc aggctgtccc cctccaaggc ctggctaata
     1441 atgtactctc aggacagact cagtatgtga ccaatgtacc agtggccctg aatgggaaca
     1501 tcaccttgct acctgtcaac agcgtttctg cagctacctt gactcccagc tctcaggcag
     1561 tcacgatcag cagctctggg tcccaggaga gtggctcaca gcctgtcacc tcagggacta
     1621 ccatcagttc tgccagcttg gtatcatcac aagccagttc cagctccttt ttcaccaatg
     1681 ccaatagcta ctcaactact actaccacca gcaacatggg aattatgaac tttactacca
     1741 gtggatcatc agggaccaac tctcaaggcc agacacccca gagggtcagt gggctacagg
     1801 ggtctgatgc tctgaacatc cagcaaaacc agacatctgg aggctcattg caagcaggcc
     1861 agcaaaaaga aggagagcaa aaccagcaga cacagcagca acaaattctt atccagcctc
     1921 agctagttca agggggacag gccctccagg ccctccaagc agcaccattg tcagggcaga
     1981 cctttacaac tcaagccatc tcccaggaaa ccctccagaa cctccagctt caggctgttc
     2041 caaactctgg tcccatcatc atccggacac caacagtggg gcccaatgga caggtcagtt
     2101 ggcagactct acagctgcag aacctccaag ttcagaaccc acaagcccaa acaatcacct
     2161 tagccccaat gcagggtgtt tccttggggc agaccagcag cagcaacacc actctcacac
     2221 ccattgcctc agctgcttcc attcctgctg gcacagtcac tgtgaatgct gctcaactct
     2281 cctccatgcc aggcctccag accattaacc tcagtgcatt gggtacttca ggaatccagg
     2341 tgcacccaat tcaaggcctg ccgttggcta tagcaaatgc cccaggtgat catggagctc
     2401 agcttggtct ccatggggct ggtggtgatg gaatacatga tgacacagca ggtggagagg
     2461 aaggagaaaa cagcccagat gcccaacccc aagccggtcg gaggacccgg cgggaagcat
     2521 gcacctgccc ctactgtaaa gacagtgaag gaaggggctc gggggatcct ggcaaaaaga
     2581 aacagcatat ttgccacatc caaggctgtg ggaaagtgta tggcaagacc tctcacctgc
     2641 gggcacactt gcgctggcat acaggcgaga ggccatttat gtgtacctgg tcatactgtg
     2701 ggaaacgctt cacacgttcg gatgagctac agaggcacaa acgtacacac acaggtgaga
     2761 agaaatttgc ctgccctgag tgtcctaagc gcttcatgag gagtgaccac ctgtcaaaac
     2821 atatcaagac ccaccagaat aagaagggag gcccaggtgt agctctgagt gtgggcactt
     2881 tgcccctgga cagtggggca ggttcagaag gcagtggcac tgccactcct tcagccctta
     2941 ttaccaccaa tatggtagcc atggaggcca tctgtccaga gggcattgcc cgtcttgcca
     3001 acagtggcat caacgtcatg caggtggcag atctgcagtc cattaatatc agtggcaatg
     3061 gcttctgaga tcaggcaccc ggggccagag acatatgggc catacccctt aaccccggga
     3121 tgcaaggtag catgggtcca agagacatgg aagagagagc catgaagcat taaaatgcat
     3181 ggtgttgaga agaatcagga gagggataca agagaggaga tggggtcccg gcacccatct
     3241 gtatcatcag tgcctctttg aaggtgggaa acattagtga aaattctgtt ggtgccacgc
     3301 tttgatgagc atttgtttga ccccagtttc ttcttacact tcttacccca gcctaccctt
     3361 cctgcatttc tcttctcagc tcttccatga tggattcccc cccctttcct aaagccatca
     3421 tgccttgata aatatatatg atcattgaaa tactttttaa caaaaaacag attctatatt
     3481 attatatata tatatatata taaagatata tagagatgca ttcacagggg ttggctggga
     3541 ggaggaagac cattctgtga ccaaaatacc ttggtcattt tttttatatt gccttatttc
     3601 cctatggctg agccttgttg tgacacatca agcttttctg tagatgttgt cttggcttcc
     3661 caccagctta agcgttcata tgctctgctt ttagttcata tatacataca taatgttttt
     3721 cctttcttaa ttttgtcttt ttgtttggga tcagcttctt gcactccttc cctaactcaa
     3781 ctgttgccgt ctcatcttct ctcatctgat cacttcatgt tttgtttttg ttactgcctg
     3841 gatgaggcac ttctgtcaat tttttcagga ccttagttcc agcagcagaa tggaaaaatc
     3901 cttgaagccc aggctgatgc ttgaagtaac tgtggaggga gtgttcaaaa tactactgac
     3961 gcaggcacct tcttggcgct ggagagtcaa aggcatctcc cttcattagc tgctctgagc
     4021 atcaagaatt agaagtcttt cagtggaatt gtacaagagt ccctttgaag ataataatct
     4081 tggctcagtt tgtataaact gtcaaatttt caaataatag gtagggggct ttcactagga
     4141 aaatcatgtg ctcagaagag gaaatgactc gtagtcaggt tcaggagtta gtggagtatt
     4201 tggactttgg tactgctgtc ttccaaggta gctctaagtt ttgatgtgtg ggcttctgag
     4261 tttatattct gaaaggaaat acacttcttt tgaacatccc cactaggttc ttttccattg
     4321 tcaataagga gcatcagcca gtgaatctgt ttcaggtttc cattctgcag aactcctcca
     4381 aagcatgtgc tagtggcaag acagtggttc ttatgatgtt ttcccttaac ttttccttgt
     4441 atgttcttgg gtggttccta agggaaaggg aagcacatga tcatgggaat gatagcccag
     4501 aacaaaaaga aatcttgtct taccacagtg ttttatagga gagattggga gaaatcatcc
     4561 tgttttctct gtgacctgat ttcagaagag actgatccaa aaattataac ggcagggaac
     4621 ctagtgcatt tggcactgag atttaaatgc aaccagaatt gtcctcaagg cccagccata
     4681 aaagcattgt ctctctcgac cttctggtat cttgttagag agcttttcac tgtgaggaag
     4741 tgtggaaaaa tagctctgtg tgtgtgtgtg tgtgtgtgtg tgtgtaatct gttaggttgg
     4801 ggataggttt tctgctagcc aatattaaaa gagacctgca ccggaattcc ctcgagctca
     4861 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tcggccgcga ctctagatca
     4921 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     4981 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     5041 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5101 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     5161 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5221 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     5281 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5341 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     5401 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5461 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5521 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5581 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5641 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5701 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     5761 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     5821 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     5881 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     5941 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     6001 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     6061 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     6121 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     6181 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     6241 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     6301 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     6361 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     6421 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     6481 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     6541 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     6601 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     6661 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     6721 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     6781 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     6841 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     6901 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     6961 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     7021 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     7081 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     7141 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     7201 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     7261 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     7321 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     7381 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7441 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7501 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     7561 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     7621 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     7681 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     7741 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     7801 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     7861 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     7921 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     7981 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8041 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8101 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8161 gctggccttt tgctca
//
