LOCUS       sequence-157203-        6230 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 6230)
  AUTHORS   Alford SC, Abdelfattah AS, Ding Y, Campbell RE
  TITLE     A fluorogenic red fluorescent protein heterodimer.
  JOURNAL   Chem Biol. 2012 Mar 23;19(3):353-60.
  PUBMED    22444590
REFERENCE   2  (bases 1 to 6230)
  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: "Chem 
            Biol."; date: "2012-03-23"; volume: "19(3)"; pages: "353-60"
COMMENT     Sequence Label: pM13-DDRFPB1
FEATURES             Location/Qualifiers
     source          1..6230
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(56..847,848..916))
                     /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     679..698
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(917..1021)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1096..1115)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1287..1666
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1667..1870
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1821..1841
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1915..1934
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1915..1933
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2237..2255)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2505..2524
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2699..2722
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2873..2890)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2879..3103
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3149..3577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3236..3255)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3446..3467
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3586..3606)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3591..3920
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3771..3906
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3833..3852
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3987..4781
                     /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(4041..4060)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4651..4670
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4955..5076
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4992..5011)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5046..5065
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5125..5141)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5125..5141)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5138..5160)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5149..5165
                     /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(5173..5179,5180..5197,5198..5203))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5218..5239
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5356..5373)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5527..6115)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5607..5626)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
       61 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
      121 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
      181 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
      241 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
      301 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
      361 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
      421 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
      481 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
      541 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
      601 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
      661 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
      721 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
      781 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
      841 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
      901 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
      961 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1021 gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct
     1081 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc
     1141 tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca
     1201 aggcttgacc gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg
     1261 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
     1321 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1381 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1441 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1501 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     1561 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     1621 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     1681 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     1741 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     1801 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     1861 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
     1921 gactcactat agggagaccc aagctggcta gcgtttaaac ttaagcttgc caccatgaag
     1981 aggcgctggc agaaaacagg ccatgctgtc cgagcattcg gaagactgaa gaagatctcc
     2041 agctccgggg cactggggca tggcaccggc agcaccggca gcggcagctc cggtaccgcc
     2101 tcctccgagg acaacaacat ggccaccatc aaagagttca tgcgcttcaa ggtgcgcatg
     2161 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     2221 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     2281 gacatcctgt ccccccagtt catgtacggc tccgaggcgt acgtgaggca ccccgccgac
     2341 atccccgatt acaagaagct gcccttcccc gagggcttca agtgggagcg cgtgatgaac
     2401 ttcgaagacg gcggtctggt gaccgttacc caggactcct ccctgcagga cggcacgctg
     2461 atctgcaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     2521 aagaccatgg gctgggaggc ctccaccgag atgctgtacc ccgaagacgg cgtgctgaag
     2581 ggccatagct atcaggccct gaagctgaag gacggcggcc actacctggt ggagttcgag
     2641 accatctaca tggccaagaa gcccgtgcaa ctgcccggcg attactgtgt ggacaccaag
     2701 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacgg gcgctccgag
     2761 ggccgccacc gtctgggcat ggacgagcgg tacaagttag ccttgaaatt agcaggtctt
     2821 gatatcggga gctaactcga gtctagaggg cccgtttaaa cccgctgatc agcctcgact
     2881 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     2941 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     3001 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     3061 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     3121 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     3181 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3241 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3301 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3361 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3421 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3481 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3541 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     3601 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3661 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3721 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     3781 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     3841 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     3901 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     3961 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4021 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4081 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4141 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4201 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4261 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4321 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4381 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     4441 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     4501 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     4561 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     4621 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     4681 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     4741 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     4801 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4861 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4921 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4981 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5041 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     5101 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5161 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5221 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5281 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     5341 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     5401 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     5461 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     5521 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5581 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5641 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5701 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5761 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5821 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5881 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5941 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6001 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6061 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6181 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
//
