LOCUS       sequence-454827-        7737 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 7737)
  TITLE     McPherson Lab unplublished
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7737)
  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: 
            "Unpublished"
COMMENT     Sequence Label: Amphiphysin Type I/Full Length
FEATURES             Location/Qualifiers
     source          1..7737
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(453..471)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(454..471)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(491..508)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    497..721
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      767..1195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(854..873)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1064..1085
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1204..1224)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1209..1538
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1389..1524
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1451..1470
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1605..2399
                     /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(1659..1678)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2269..2288
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2573..2694
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2610..2629)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2664..2683
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2743..2759)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2743..2759)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2756..2778)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2767..2783
                     /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(2791..2797,2798..2815,2816..2821))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2836..2857
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2974..2991)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3145..3733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3225..3244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3904..4695,4696..4764))
                     /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     4527..4546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4765..4869)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4944..4963)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5135..5514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5515..5718
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5669..5689
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5763..5782
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5763..5781
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aaggtggaaa cactgcatga ttttgaggca gcaaattctg atgaacttac cttacaaagg
       61 ggtgatgtgg tgctggtggt cccctcagat tcagaagctg atcaggatgc aggctggctg
      121 gtgggagtga aggaatcaga ctggcttcag tacagagacc ttgccaccta caaaggcctc
      181 tttccagaga acttcacccg acgcttagat tagggcaaca agtactgcaa gaaggagctc
      241 agttacgggg tttttaaacc ttcatgaaaa cctgaagagt tcacttttgt tattatgctc
      301 ttaatgattt acagactgat gccagacaaa ccttgggaag atgtatcaat ggagcatgtg
      361 tgcaaaaaaa tgtaagagga aaaaaaaaac cggaattctg cagatatcca tcacactggc
      421 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
      481 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
      541 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
      601 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
      661 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
      721 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
      781 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      841 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      901 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      961 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1021 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1081 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1141 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     1201 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     1261 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     1321 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     1381 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     1441 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     1501 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     1561 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     1621 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     1681 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     1741 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     1801 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     1861 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     1921 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     1981 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     2041 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     2101 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     2161 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     2221 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     2281 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     2341 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     2401 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     2461 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     2521 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     2581 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2641 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     2701 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     2761 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     2821 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     2881 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2941 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3001 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3061 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     3181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     3241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     3301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     3361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     3421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     3481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     3541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     3901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     3961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     4201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     4261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     4321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     4381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     4441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     4501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     4561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     4621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     4681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     4741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     4801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     4861 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     4921 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     4981 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     5041 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     5101 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     5161 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     5221 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     5281 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     5341 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     5401 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     5461 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     5521 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     5581 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     5641 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     5701 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     5761 attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat ccactagtaa
     5821 cggccgccag tgtgctggaa ttccgggccc cccggacccg cagccatggc cgacatcaag
     5881 acgggcatct tcgccaagaa cgtccagaag cgactcaacc gcgcgcagga aaaggtcctc
     5941 caaaagctgg ggaaagctga tgagacaaaa gacgaacagt tcgaagaata tgtccagaac
     6001 ttcaaacggc aagaagcaga gggtaccaga cttcagcgag aactccgagg atatttagca
     6061 atcaaaggca tgcaggaggc ctccatgaag ctcacagagt cgctgcatga agtctatgag
     6121 cctgactggt atgggcggga agatgtgaaa atggttggtg agaaatgtga tgtgctgtgg
     6181 gaagacttcc atcaaaaact cgtggatggg tccttgctaa cactggatac ctacctgggg
     6241 caatttcctg acataaagaa tcgcatcgcc aagcgcagca ggaagctagt ggactatgac
     6301 agtgcccgcc accatctgga agctctgcag agctccaaga ggaaggatga gagtcgaatc
     6361 tctaaggcag aagaagaatt tcagaaagca cagaaagtgt ttgaagagtt taacgttgac
     6421 ttacaagaag agttaccatc attatggtca agacgagttg gattttatgt taatactttc
     6481 aaaaacgtct ccagccttga agccaagttt cataaggaaa ttgcggtgct ttgccacaaa
     6541 ctgtatgaag tgatgacaaa actgggtgac cagcacgccg acaaggcctt caccatccaa
     6601 ggagcgccca gtgattcggg tcctctccgc attgcaaaga caccatcacc gcctgaggag
     6661 ccttcacccc tcccgagccc gacagcaagt ccaaatcata cattagcacc tgcgtctccc
     6721 gcaccagcac ggcctcggtc accttcacag acaaggaaag ggcctcctgt cccacctcta
     6781 cctaaagtca ccccgacaaa ggaactgcag caggagaaca tcatcagttt ctttgaggac
     6841 aactttgttc cagaaatcag tgtgacaaca ccttcccaga atgaagtccc tgaggtgaag
     6901 aaagaggaga ctttgctgga tctggacttt gatcctttca agcccgaggt gacacctgca
     6961 ggttctgctg gagtgaccca ctcacccatg tctcagacat tgccctggga cctatggacg
     7021 acaagcactg atttggtaca gccggcttct ggtggttcat ttaatggatt cacacagccc
     7081 caggatactt cattattcac aatgcagaca gaccagagta tgatctgcaa cttggctgaa
     7141 tctgaacagg ctccacccac agagccaaaa gcagaggagc ctctggctgc tgtcacacct
     7201 gccgttggtc tggaccttgg aatggacact cgggctgagg agccagtgga ggaggcagtg
     7261 atcatacctg gagctgatgc tgatgcagct gttggaacct tggtgtcagc agctgagggg
     7321 gccccaggag aggaagcaga ggcggagaag gccactgtcc ctgccgggga aggagtaagt
     7381 ttagaggagg ccaaaattgg aactgaaacc actgagggtg cagagagtgc ccaacctgaa
     7441 gcagaggagc tcgaagcaac agtgcctcag gagaaggtca ttccttcggt ggtcatagag
     7501 cctgcctcca accatgaaga ggaaggagaa aacgaaataa ctataggtgc agagcccaag
     7561 gagaccaccg aggacgcggc tcctccgggc cccaccagcg agacaccgga gctggctacg
     7621 gagcagaagc ctatccagga ccctcagccc acgccttctg caccagccat gggggctgct
     7681 gaccagctag catctgcaag ggaggcctct caggaattgc ctcctggctt tctctac
//
