LOCUS       sequence-8892-ex        4139 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 4139)
  AUTHORS   Groth AC, Olivares EC, Thyagarajan B, Calos MP
  TITLE     A phage integrase directs efficient site-specific integration in 
            human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2000 May 23. 97(11):5995-6000.
  PUBMED    10801973
REFERENCE   2  (bases 1 to 4139)
  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: "Proc Natl 
            Acad Sci U S A. 2000 May 23. 97(11):5995-6000."
COMMENT     Sequence Label: pTA-attP
FEATURES             Location/Qualifiers
     source          1..4139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    72..171
                     /label=phage phi-C31 attP
                     /bound_moiety="phage phi-C31 integrase"
                     /note="attachment site of phage phi-C31"
     primer_bind     complement(302..321)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(303..321)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(328..345)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(328..344)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(337..359)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      486..866
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(573..592)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     783..804
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             1234..2028
                     /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
                     SRLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVIHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1288..1307)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             join(2046..2114,2115..2906)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDESDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3077..3665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3566..3585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3819..3836
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3953..3974
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3989..3994,3995..4012,4013..4019)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4027..4043
                     /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)."
     primer_bind     4032..4054
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4051..4067
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4051..4067
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgactagtac tgacggacac accgaagccc cggcggcaac cctcagcgga tgccccgggg
       61 cttcacgttt tcccaggtca gaagcggttt tcgggagtag tgccccaact ggggtaacct
      121 ttgagttctc tcagttgggg gcgtagggtc gccgacatga cacaaggggt tgtgaccggg
      181 gtggacacgt acgcgggtgc ttacgaccgt cagtcgcgcg agcgcgacta gtacagccga
      241 attctgcaga tatccatcac actggcggcc gctcgagcat gcatctagag ggcccaattc
      301 gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc gtgactggga
      361 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
      421 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
      481 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
      541 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
      601 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
      661 atttagagct ttacggcacc tcgaccgcaa aaaacttgat ttgggtgatg gttcacgtag
      721 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
      781 tagtggactc ttgttccaaa ctggaacaac actcaaccct atcgcggtct attcttttga
      841 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaat
      901 tcagggcgca agggctgcta aaggaaccgg aacacgtaga aagccagtcc gcagaaacgg
      961 tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa cgcaagcgca
     1021 aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg ggcggtttta
     1081 tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt tgggaagccc
     1141 tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag gggatcaaga
     1201 tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     1261 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     1321 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     1381 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     1441 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     1501 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctcg ccttgctcct
     1561 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     1621 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     1681 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     1741 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gatccatggc
     1801 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt caacgactgt
     1861 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggatacccg tgatattgct
     1921 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     1981 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga
     2041 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2101 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2161 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2221 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt catacactat
     2281 tatcccgtat tgacgccggg caagagcaac tcggtcgccg ggcgcggtat tctcagaatg
     2341 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2401 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     2461 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     2521 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag agtgacacca
     2581 cgatgcctgt agcaatgcca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     2641 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     2701 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     2761 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     2821 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     2881 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     2941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     3241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg
     3481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     3541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     3601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     3661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     3721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     3781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     3841 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     3901 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     3961 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     4021 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     4081 agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga attcggctt
//
